A high-content image-based drug screen of clinical compounds against cell transmission of adenovirus

<!DOCTYPE html> <html lang="en" > <head > <meta charset="UTF-8" /> <meta http-equiv="X-UA-Compatible" content="IE=edge" /> <meta name="HandheldFriendly" content="True" /> <meta name="MobileOptimized" content="320" /> <meta name="viewport" content="width=device-width, initial-scale=1.0" /> <link rel="stylesheet" href="/static/assets/style-70b9163a.css" /> <script type="module" crossorigin="" src="/static/assets/base_style-ec2bc71e.js"></script> <link rel="stylesheet" href="/static/assets/style-ef962842.css" /> <link rel="stylesheet" href="/static/assets/style-3ade8b5c.css" /> <script type="module" crossorigin="" src="/static/assets/article_style-d757a0dd.js"></script> <style> @media screen and (min-width: 64em) { div.pmc-wm { background: repeat-y; background-image: url("data:image/svg+xml,%3Csvg xmlns='http://www.w3.org/2000/svg' width='20' height='350' xmlns:xlink='http://www.w3.org/1999/xlink'%3E%3Cdefs%3E%3Cfilter x='-.02' y='0' width='1.05' height='1' id='c'%3E%3CfeFlood flood-color='%23FFF'/%3E%3CfeComposite in='SourceGraphic'/%3E%3C/filter%3E%3Ctext id='b' font-family='Helvetica' font-size='11pt' style='opacity:1;fill:%23005ea2;stroke:none;text-anchor:middle' x='175' y='14'%3E%3C/text%3E%3Cpath id='a' style='fill:%23005ea2' d='M0 8h350v3H0z'/%3E%3C/defs%3E%3Cuse xlink:href='%23a' transform='rotate(90 10 10)'/%3E%3Cuse xlink:href='%23b' transform='rotate(90 10 10)' filter='url(%23c)'/%3E%3C/svg%3E"); padding-left: 3rem; } } </style> <link rel="apple-touch-icon" sizes="180x180" href="/static/img/favicons/apple-touch-icon.png" /> <link rel="icon" type="image/png" sizes="48x48" href="/static/img/favicons/favicon-48x48.png" /> <link rel="icon" type="image/png" sizes="32x32" href="/static/img/favicons/favicon-32x32.png" /> <link rel="icon" type="image/png" sizes="16x16" href="/static/img/favicons/favicon-16x16.png" /> <link rel="manifest" href="/static/img/favicons/site.webmanifest" /> <link rel="mask-icon" href="/static/img/favicons/safari-pinned-tab.svg" color="#0071bc" /> <meta name="msapplication-config" content="/static/img/favicons/browserconfig.xml" /> <meta name="theme-color" content="#ffffff" /> <title> A high-content image-based drug screen of clinical compounds against cell transmission of adenovirus - PMC </title>  <meta name="ncbi_app" content="cloudpmc-viewer" /> <meta name="ncbi_db" content="pmc" /> <meta name="ncbi_phid" content="6E4B00557471BEF30600550000AE8103.m_1" />  <meta name="ncbi_pdid" content="article" /> <link rel="preconnect" href="https://www.google-analytics.com" /> <link rel="dns-prefetch" href="https://cdn.ncbi.nlm.nih.gov" /> <link rel="preconnect" href="https://code.jquery.com" /> <meta name="ncbi_domain" content="sdata"> <meta name="ncbi_type" content="fulltext"> <meta name="ncbi_pcid" content="journal"> <meta name="ncbi_feature" content="associated_data"> <link rel="canonical" href="https://pmc.ncbi.nlm.nih.gov/articles/PMC7423605/"> <meta name="robots" content="INDEX,NOFOLLOW,NOARCHIVE"> <meta name="citation_journal_title" content="Scientific Data"> <meta name="citation_title" content="A high-content image-based drug screen of clinical compounds against cell transmission of adenovirus"> <meta name="citation_author" content="Fanny Georgi"> <meta name="citation_author_institution" content="Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland"> <meta name="citation_author" content="Fabien Kuttler"> <meta name="citation_author_institution" content="Biomolecular Screening Facility, School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL), Station 15, Lausanne, 1015 Switzerland"> <meta name="citation_author" content="Luca Murer"> <meta name="citation_author_institution" content="Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland"> <meta name="citation_author" content="Vardan Andriasyan"> <meta name="citation_author_institution" content="Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland"> <meta name="citation_author" content="Robert Witte"> <meta name="citation_author_institution" content="Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland"> <meta name="citation_author" content="Artur Yakimovich"> <meta name="citation_author_institution" content="MRC Laboratory for Molecular Cell Biology, University College London, Gower St, London, WC1E 6BT United Kingdom"> <meta name="citation_author_institution" content="Artificial Intelligence for Life Sciences CIC, 40 Gowers walk, London, E1 8BH United Kingdom"> <meta name="citation_author" content="Gerardo Turcatti"> <meta name="citation_author_institution" content="Biomolecular Screening Facility, School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL), Station 15, Lausanne, 1015 Switzerland"> <meta name="citation_author" content="Urs F Greber"> <meta name="citation_author_institution" content="Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland"> <meta name="citation_publication_date" content="2020 Aug 12"> <meta name="citation_volume" content="7"> <meta name="citation_firstpage" content="265"> <meta name="citation_doi" content="10.1038/s41597-020-00604-0"> <meta name="citation_pmid" content="32788590"> <meta name="citation_abstract_html_url" content="https://pmc.ncbi.nlm.nih.gov/articles/PMC7423605/"> <meta name="citation_fulltext_html_url" content="https://pmc.ncbi.nlm.nih.gov/articles/PMC7423605/"> <meta name="citation_pdf_url" content="https://pmc.ncbi.nlm.nih.gov/articles/PMC7423605/pdf/41597_2020_Article_604.pdf"> <meta name="description" content="Human adenoviruses (HAdVs) are fatal to immuno-suppressed individuals, but no effective anti-HAdV therapy is available. Here, we present a novel image-based high-throughput screening (HTS) platform, which scores the full viral replication cycle from ..."> <meta name="og:title" content="A high-content image-based drug screen of clinical compounds against cell transmission of adenovirus"> <meta name="og:type" content="article"> <meta name="og:site_name" content="PubMed Central (PMC)"> <meta name="og:description" content="Human adenoviruses (HAdVs) are fatal to immuno-suppressed individuals, but no effective anti-HAdV therapy is available. Here, we present a novel image-based high-throughput screening (HTS) platform, which scores the full viral replication cycle from ..."> <meta name="og:url" content="https://pmc.ncbi.nlm.nih.gov/articles/PMC7423605/"> <meta name="og:image" content="https://cdn.ncbi.nlm.nih.gov/pmc/cms/images/pmc-card-share.jpg?_=0"> <meta name="twitter:card" content="summary_large_image"> <meta name="twitter:site" content="@ncbi"> </head> <body > <a class="usa-skipnav " href="#main-content"> Skip to main content </a> <section class="usa-banner " aria-label="Official website of the United States government" > <div class="usa-accordion"> <header class="usa-banner__header"> <div class="usa-banner__inner"> <div class="grid-col-auto"> <img aria-hidden="true" class="usa-banner__header-flag" src="/static/img/us_flag.svg" alt="" /> </div> <div class="grid-col-fill tablet:grid-col-auto" aria-hidden="true"> <p class="usa-banner__header-text"> An official website of the United States government </p> <span class="usa-banner__header-action">Here's how you know</span> </div> <button type="button" class="usa-accordion__button usa-banner__button " aria-expanded="false" aria-controls="gov-banner-default" data-testid="storybook-django-banner" > <span class="usa-banner__button-text">Here's how you know</span> </button> </div> </header> <div class="usa-banner__content usa-accordion__content" id="gov-banner-default" hidden> <div class="grid-row grid-gap-lg"> <div class="usa-banner__guidance tablet:grid-col-6"> <img class="usa-banner__icon usa-media-block__img" src="/static/img/icon-dot-gov.svg" alt="" aria-hidden="true" /> <div class="usa-media-block__body"> <p> <strong>Official websites use .gov</strong> <br /> A <strong>.gov</strong> website belongs to an official government organization in the United States. </p> </div> </div> <div class="usa-banner__guidance tablet:grid-col-6"> <img class="usa-banner__icon usa-media-block__img" src="/static/img/icon-https.svg" alt="" aria-hidden="true" /> <div class="usa-media-block__body"> <p> <strong>Secure .gov websites use HTTPS</strong> <br /> A <strong>lock</strong> ( <span class="icon-lock"> <svg xmlns="http://www.w3.org/2000/svg" width="52" height="64" viewBox="0 0 52 64" class="usa-banner__lock-image" role="graphics-symbol" aria-labelledby="banner-lock-description" focusable="false"> <title id="banner-lock-title">Lock</title> <desc id="banner-lock-description"> Locked padlock icon </desc> <path fill="#000000" fill-rule="evenodd" d="M26 0c10.493 0 19 8.507 19 19v9h3a4 4 0 0 1 4 4v28a4 4 0 0 1-4 4H4a4 4 0 0 1-4-4V32a4 4 0 0 1 4-4h3v-9C7 8.507 15.507 0 26 0zm0 8c-5.979 0-10.843 4.77-10.996 10.712L15 19v9h22v-9c0-6.075-4.925-11-11-11z" /> </svg> </span>) or <strong>https://</strong> means you've safely connected to the .gov website. Share sensitive information only on official, secure websites. </p> </div> </div> </div> </div> </div> </section> <div class="usa-overlay"> </div> <header class="usa-header usa-header--extended usa-header--wide" data-testid="header" data-header > <div class="ncbi-header"> <div class="ncbi-header__container"> <a class="ncbi-header__logo-container" href="/"> <img alt=" PMC home page " class="ncbi-header__logo-image" src="/static/img/ncbi-logos/nih-nlm-ncbi--white.svg" /> </a>  <button type="button" class="usa-menu-btn ncbi-header__hamburger-button " aria-label="Show menu" data-testid="navMenuButton" > <svg aria-hidden="true" class="ncbi-hamburger-icon" fill="none" focusable="false" height="21" viewBox="0 0 31 21" width="31" xmlns="http://www.w3.org/2000/svg"> <path clip-rule="evenodd" d="M0.125 20.75H30.875V17.3333H0.125V20.75ZM0.125 12.2083H30.875V8.79167H0.125V12.2083ZM0.125 0.25V3.66667H30.875V0.25H0.125Z" fill="#F1F1F1" fill-rule="evenodd" /> </svg> </button>  <div class="ncbi-header__desktop-buttons">  <button type="button" class="usa-button usa-button--unstyled ncbi-header__desktop-button " aria-expanded="false" aria-controls="search-field-desktop-navigation" aria-label="Show search overlay" data-testid="toggleSearchPanelButton" data-toggle-search-panel-button > <svg class="usa-icon " role="graphics-symbol" aria-hidden="true" > <use xlink:href="/static/img/sprite.svg#search" /> </svg> Search </button>  <div class="ncbi-header__login-dropdown"> <button type="button" class="usa-button usa-button--unstyled ncbi-header__desktop-button ncbi-header__login-dropdown-button " aria-expanded="false" aria-controls="login-dropdown-menu" aria-label="Show login menu" data-testid="toggleLoginMenuDropdown" data-desktop-login-button > <svg class="usa-icon " role="graphics-symbol" aria-hidden="true" > <use xlink:href="/static/img/sprite.svg#person" /> </svg> <span data-login-dropdown-text>Log in</span>  <svg class="usa-icon ncbi-header__login-dropdown-icon ncbi-header__login-dropdown-icon--expand-less ncbi-header__login-dropdown-icon--hidden" role="graphics-symbol" aria-hidden="true" data-login-dropdown-up-arrow> <use xlink:href="/static/img/sprite.svg#expand_less" /> </svg>  <svg class="usa-icon ncbi-header__login-dropdown-icon ncbi-header__login-dropdown-icon--expand-more ncbi-header__login-dropdown-icon--hidden" role="graphics-symbol" aria-hidden="true" data-login-dropdown-down-arrow> <use xlink:href="/static/img/sprite.svg#expand_more" /> </svg> </button>  <ul class="usa-nav__submenu ncbi-header__login-dropdown-menu" id="login-dropdown-menu" data-desktop-login-menu-dropdown hidden> <li class="usa-nav__submenu-item">  <a href="https://www.ncbi.nlm.nih.gov/myncbi/" class="usa-link " > Dashboard </a> </li> <li class="usa-nav__submenu-item">  <a href="https://www.ncbi.nlm.nih.gov/myncbi/collections/bibliography/" class="usa-link " > Publications </a> </li> <li class="usa-nav__submenu-item">  <a href="https://www.ncbi.nlm.nih.gov/account/settings/" class="usa-link " > Account settings </a> </li> <li class="usa-nav__submenu-item"> <button type="button" class="usa-button usa-button--outline ncbi-header__login-dropdown-logout-button " data-testid="desktopLogoutButton" data-desktop-logout-button > Log out </button> </li> </ul> </div> </div> </div> </div>  <div class="ncbi-search-panel ncbi--show-only-at-desktop" data-testid="searchPanel" data-header-search-panel hidden> <div class="ncbi-search-panel__container"> <form action="https://www.ncbi.nlm.nih.gov/search/all/" aria-describedby="search-field-desktop-navigation-help-text" autocomplete="off" class="usa-search usa-search--big ncbi-search-panel__form" data-testid="form" method="GET" role="search"> <label class="usa-sr-only" data-testid="label" for="search-field-desktop-navigation"> Search… </label> <input class="usa-input" data-testid="textInput" id="search-field-desktop-navigation" name="term" placeholder="Search NCBI" type="search" value="" /> <button type="submit" class="usa-button " data-testid="button" > <span class="usa-search__submit-text"> Search NCBI </span> </button> </form> </div> </div> <nav aria-label="Primary navigation" class="usa-nav"> <p class="usa-sr-only" id="primary-navigation-sr-only-title"> Primary site navigation </p>  <button type="button" class="usa-nav__close ncbi-nav__close-button " aria-label="Close navigation menu" data-testid="navCloseButton" > <img src="/static/img/usa-icons/close.svg" alt="Close" /> </button>  <form class="usa-search usa-search--small ncbi--hide-at-desktop margin-top-6" role="search"> <label class="usa-sr-only" for="search-field"> Search </label> <input class="usa-input" id="search-field-mobile-navigation" type="search" placeholder="Search NCBI" name="search" /> <button type="submit" class="usa-button " >  <img src="data:image/svg+xml;base64,PHN2ZyB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciIGhlaWdodD0iMjQiIHZpZXdCb3g9IjAgMCAyNCAyNCIgd2lkdGg9IjI0Ij48cGF0aCBkPSJNMCAwaDI0djI0SDB6IiBmaWxsPSJub25lIi8+PHBhdGggZmlsbD0iI2ZmZiIgZD0iTTE1LjUgMTRoLS43OWwtLjI4LS4yN0E2LjQ3MSA2LjQ3MSAwIDAgMCAxNiA5LjUgNi41IDYuNSAwIDEgMCA5LjUgMTZjMS42MSAwIDMuMDktLjU5IDQuMjMtMS41N2wuMjcuMjh2Ljc5bDUgNC45OUwyMC40OSAxOWwtNC45OS01em0tNiAwQzcuMDEgMTQgNSAxMS45OSA1IDkuNVM3LjAxIDUgOS41IDUgMTQgNy4wMSAxNCA5LjUgMTEuOTkgMTQgOS41IDE0eiIvPjwvc3ZnPg==" class="usa-search__submit-icon" alt="Search" /> </button> </form>   <div class="ncbi-nav__mobile-login-menu ncbi--hide-at-desktop" data-mobile-login-menu hidden> <p class="ncbi-nav__mobile-login-menu-status"> Logged in as: <strong class="ncbi-nav__mobile-login-menu-email" data-mobile-login-email-text></strong> </p> <ul class="usa-nav__primary usa-accordion"> <li class="usa-nav__primary-item"> <a href="https://www.ncbi.nlm.nih.gov/myncbi/" class="usa-link " > Dashboard </a> </li> <li class="usa-nav__primary-item"> <a href="https://www.ncbi.nlm.nih.gov/myncbi/collections/bibliography/" class="usa-link " > Publications </a> </li> <li class="usa-nav__primary-item"> <a href="https://www.ncbi.nlm.nih.gov/account/settings/" class="usa-link " > Account settings </a> </li> </ul> </div> <button type="button" class="usa-button ncbi-nav__mobile-login-button ncbi--hide-at-desktop " data-testid="mobileLoginButton" data-mobile-login-button > Log in </button> </nav> </header> <section class="pmc-header pmc-header--basic" aria-label="PMC Header with search box"> <div class="pmc-nav-container"> <div class="pmc-header__bar"> <div class="pmc-header__logo"> <a href="/" title="Home" aria-label="PMC Home"></a> </div> <button type="button" class="usa-button usa-button--unstyled pmc-header__search__button" aria-label="Open search" data-ga-category="search" data-ga-action="PMC" data-ga-label="pmc_search_panel_mobile" > <svg class="usa-icon width-4 height-4 pmc-icon__open" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#search"></use> </svg> <svg class="usa-icon width-4 height-4 pmc-icon__close" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#close"></use> </svg> </button> </div> <div class="pmc-header__search"> <form class="usa-search usa-search--extra usa-search--article-right-column pmc-header__search__form" autocomplete="off" role="search"> <label class="usa-sr-only" for="pmc-search">Search PMC Full-Text Archive</label> <span class="autoComplete_wrapper flex-1"> <input class="usa-input width-full maxw-none" required="required" placeholder="Search PMC Full-Text Archive" id="pmc-search" type="search" name="term" data-autocomplete-url="/search/autocomplete/"/> </span> <button class="usa-button" type="submit" formaction="https://www.ncbi.nlm.nih.gov/pmc/" data-ga-category="search" data-ga-action="PMC" data-ga-label="PMC_search_button" > <span class="usa-search__submit-text">Search in PMC</span> <img src="/static/img/usa-icons-bg/search--white.svg" class="usa-search__submit-icon" alt="Search" /> </button> </form> <ul class="pmc-header__search__menu"> <li> <a class="usa-link" href="https://www.ncbi.nlm.nih.gov/pmc/advanced/" data-ga-action="featured_link" data-ga-label="advanced_search"> Advanced Search </a> </li> <li> <a class="usa-link" href="/journals/" data-ga-action="featured_link" data-ga-label="journal list"> Journal List </a> </li> <li> <a class="usa-link" href="/about/userguide/" data-ga-action="featured_link" data-ga-label="user guide"> User Guide </a> </li> </ul> </div> </div> </section> <div class="usa-section padding-top-0 desktop:padding-top-6 pmc-article-section" data-article-db="pmc" data-article-id="7423605"> <div class="grid-container pmc-actions-bar" aria-label="Actions bar" role="complementary"> <div class="grid-row"> <div class="grid-col-fill display-flex"> <div class="display-flex"> <ul class="usa-list usa-list--unstyled usa-list--horizontal"> <li class="margin-right-2 mobile-lg:margin-right-4 display-flex mob"> <button type="button" class="usa-button pmc-sidenav__container__open usa-button--unstyled width-auto display-flex" aria-label="Open resources" data-extra-class="is-visible-resources" data-ga-category="resources_accordion" data-ga-action="click" data-ga-label="mobile_icon" > <svg class="usa-icon width-4 height-4" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#more_vert"></use> </svg> </button> </li> <li class="margin-right-2 mobile-lg:margin-right-4 display-flex mob"> <a href="https://doi.org/10.1038/s41597-020-00604-0" class="usa-link display-flex" role="button" target="_blank" rel="noreferrer noopener" aria-label="View on publisher site" data-ga-category="actions" data-ga-action="click" data-ga-label="publisher_link_mobile" > <svg class="usa-icon width-4 height-4" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#launch"></use> </svg> </a> </li> <li class="margin-right-2 mobile-lg:margin-right-4 display-flex"> <a href="pdf/41597_2020_Article_604.pdf" class="usa-link display-flex" role="button" aria-label="Download PDF" data-ga-category="actions" data-ga-action="click" data-ga-label="pdf_download_mobile" > <svg class="usa-icon width-4 height-4" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#file_download"></use> </svg> </a> </li> <li class="margin-right-2 mobile-lg:margin-right-4 display-flex"> <button class="usa-button usa-button--unstyled collections-dialog-trigger collections-button display-flex collections-button-empty" aria-label="Save article in MyNCBI collections." data-ga-category="actions" data-ga-action="click" data-ga-label="collections_button_mobile" data-collections-open-dialog-enabled="false" data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpmc.ncbi.nlm.nih.gov%2Farticles%2FPMC7423605%2F%23open-collections-dialog" data-in-collections="false" > <svg class="usa-icon width-4 height-4 usa-icon--bookmark-full" aria-hidden="true" focusable="false" role="img" hidden> <use xlink:href="/static/img/action-bookmark-full.svg#icon"></use> </svg> <svg class="usa-icon width-4 height-4 usa-icon--bookmark-empty" aria-hidden="true" focusable="false" role="img" hidden> <use xlink:href="/static/img/action-bookmark-empty.svg#icon"></use> </svg> </button> </li> <li class="margin-right-2 mobile-lg:margin-right-4 display-flex"> <button role="button" class="usa-button usa-button--unstyled citation-dialog-trigger display-flex" aria-label="Open dialog with citation text in different styles" data-ga-category="actions" data-ga-action="open" data-ga-label="cite_mobile" data-all-citations-url="/resources/citations/7423605/" data-citation-style="nlm" data-download-format-link="/resources/citations/7423605/export/" > <svg class="usa-icon width-4 height-4 usa-icon--bookmark-empty" aria-hidden="true" focusable="false" role="img" hidden> <use xlink:href="/static/img/sprite.svg#format_quote"></use> </svg> </button> </li> <li class="pmc-permalink display-flex"> <button type="button" class="usa-button usa-button--unstyled display-flex" aria-label="Show article permalink" aria-expanded="false" aria-haspopup="true" data-ga-category="actions" data-ga-action="open" data-ga-label="permalink_mobile" > <svg class="usa-icon width-4 height-4" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#share"></use> </svg> </button> <div class="pmc-permalink__dropdown" hidden> <div class="pmc-permalink__dropdown__container"> <h2 class="usa-modal__heading margin-top-0 margin-bottom-2">PERMALINK</h2> <div class="pmc-permalink__dropdown__content"> <input type="text" class="usa-input" value="https://pmc.ncbi.nlm.nih.gov/articles/PMC7423605/" aria-label="Article permalink"> <button class="usa-button display-inline-flex pmc-permalink__dropdown__copy__btn margin-right-0" title="Copy article permalink" data-ga-category="save_share" data-ga-action="link" data-ga-label="copy_link"> <svg class="usa-icon" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#content_copy"></use> </svg> <span class="margin-left-1">Copy</span> </button> </div> </div> </div> </li> </ul> </div> <button type="button" class="usa-button pmc-sidenav__container__open usa-button--unstyled width-auto display-flex" aria-label="Open article navigation" data-extra-class="is-visible-in-page" data-ga-category="actions" data-ga-action="open" data-ga-label="article_nav_mobile" > <svg class="usa-icon width-4 height-4" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#list"></use> </svg> </button> </div> </div> </div> <div class="grid-container desktop:padding-left-6"> <div id="article-container" class="grid-row grid-gap"> <div class="grid-col-12 desktop:grid-col-8 order-2 pmc-layout__content"> <div class="grid-container padding-left-0 padding-right-0"> <div class="grid-row desktop:margin-left-neg-6"> <div class="grid-col-12"> <div class="pmc-layout__disclaimer" role="complementary" aria-label="Disclaimer note"> As a library, NLM provides access to scientific literature. Inclusion in an NLM database does not imply endorsement of, or agreement with, the contents by NLM or the National Institutes of Health.<br/> Learn more: <a class="usa-link" data-ga-category="Link click" data-ga-action="Disclaimer" data-ga-label="New disclaimer box" href="/about/disclaimer/">PMC Disclaimer</a> | <a class="usa-link" data-ga-category="Link click" data-ga-action="PMC Copyright Notice" data-ga-label="New disclaimer box" href="/about/copyright/"> PMC Copyright Notice </a> </div> </div> </div> <div class="grid-row pmc-wm desktop:margin-left-neg-6">  <main id="main-content" class="usa-layout-docs__main usa-layout-docs grid-col-12 pmc-layout pmc-prose padding-0" > <section class="pmc-journal-banner text-center line-height-none" aria-label="Journal banner"><img src="https://cdn.ncbi.nlm.nih.gov/pmc/banners/logo-sdata.jpg" alt="Scientific Data logo" usemap="#pmc-banner-imagemap" width="500" height="75"><map name="pmc-banner-imagemap"><area alt="Link to Scientific Data" title="Link to Scientific Data" shape="default" href="http://www.nature.com/sdata/" target="_blank" rel="noopener noreferrer"></map></section><article lang="en"><section aria-label="Article citation and metadata"><section class="pmc-layout__citation font-secondary font-xs"><div> <div class="display-inline-block"><button type="button" class="cursor-pointer text-no-underline bg-transparent border-0 padding-0 text-left margin-0 text-normal text-primary" aria-controls="journal_context_menu">Sci Data</button></div>. 2020 Aug 12;7:265. doi: <a href="https://doi.org/10.1038/s41597-020-00604-0" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">10.1038/s41597-020-00604-0</a> </div> <nav id="journal_context_menu" hidden="hidden"><ul class="menu-list font-family-ui" role="menu"> <li role="presentation"><a href="https://www.ncbi.nlm.nih.gov/pmc/?term=%22Sci%20Data%22%5Bjour%5D" class="usa-link" role="menuitem">Search in PMC</a></li> <li role="presentation"><a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Sci%20Data%22%5Bjour%5D" lang="en" class="usa-link" role="menuitem">Search in PubMed</a></li> <li role="presentation"><a href="https://www.ncbi.nlm.nih.gov/nlmcatalog?term=%22Sci%20Data%22%5BTitle%20Abbreviation%5D" class="usa-link" role="menuitem">View in NLM Catalog</a></li> <li role="presentation"><a href="?term=%22Sci%20Data%22%5Bjour%5D" class="usa-link" role="menuitem" data-add-to-search="true">Add to search</a></li> </ul></nav></section><section class="front-matter"><div class="ameta p font-secondary font-xs"> <hgroup><h1>A high-content image-based drug screen of clinical compounds against cell transmission of adenovirus</h1></hgroup><div class="cg p"> <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Georgi%20F%22%5BAuthor%5D" class="usa-link" aria-describedby="id1"><span class="name western">Fanny Georgi</span></a><div hidden="hidden" id="id1"> <h3><span class="name western">Fanny Georgi</span></h3> <div class="p"> <sup>1</sup>Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland </div> <div class="p">Find articles by <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Georgi%20F%22%5BAuthor%5D" class="usa-link"><span class="name western">Fanny Georgi</span></a> </div> </div> <sup>1</sup>, <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Kuttler%20F%22%5BAuthor%5D" class="usa-link" aria-describedby="id2"><span class="name western">Fabien Kuttler</span></a><div hidden="hidden" id="id2"> <h3><span class="name western">Fabien Kuttler</span></h3> <div class="p"> <sup>2</sup>Biomolecular Screening Facility, School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL), Station 15, Lausanne, 1015 Switzerland </div> <div class="p">Find articles by <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Kuttler%20F%22%5BAuthor%5D" class="usa-link"><span class="name western">Fabien Kuttler</span></a> </div> </div> <sup>2</sup>, <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Murer%20L%22%5BAuthor%5D" class="usa-link" aria-describedby="id3"><span class="name western">Luca Murer</span></a><div hidden="hidden" id="id3"> <h3><span class="name western">Luca Murer</span></h3> <div class="p"> <sup>1</sup>Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland </div> <div class="p">Find articles by <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Murer%20L%22%5BAuthor%5D" class="usa-link"><span class="name western">Luca Murer</span></a> </div> </div> <sup>1</sup>, <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Andriasyan%20V%22%5BAuthor%5D" class="usa-link" aria-describedby="id4"><span class="name western">Vardan Andriasyan</span></a><div hidden="hidden" id="id4"> <h3><span class="name western">Vardan Andriasyan</span></h3> <div class="p"> <sup>1</sup>Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland </div> <div class="p">Find articles by <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Andriasyan%20V%22%5BAuthor%5D" class="usa-link"><span class="name western">Vardan Andriasyan</span></a> </div> </div> <sup>1</sup>, <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Witte%20R%22%5BAuthor%5D" class="usa-link" aria-describedby="id5"><span class="name western">Robert Witte</span></a><div hidden="hidden" id="id5"> <h3><span class="name western">Robert Witte</span></h3> <div class="p"> <sup>1</sup>Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland </div> <div class="p">Find articles by <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Witte%20R%22%5BAuthor%5D" class="usa-link"><span class="name western">Robert Witte</span></a> </div> </div> <sup>1</sup>, <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Yakimovich%20A%22%5BAuthor%5D" class="usa-link" aria-describedby="id6"><span class="name western">Artur Yakimovich</span></a><div hidden="hidden" id="id6"> <h3><span class="name western">Artur Yakimovich</span></h3> <div class="p"> <sup>3</sup>MRC Laboratory for Molecular Cell Biology, University College London, Gower St, London, WC1E 6BT United Kingdom </div> <div class="p"> <sup>4</sup>Artificial Intelligence for Life Sciences CIC, 40 Gowers walk, London, E1 8BH United Kingdom </div> <div class="p">Find articles by <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Yakimovich%20A%22%5BAuthor%5D" class="usa-link"><span class="name western">Artur Yakimovich</span></a> </div> </div> <sup>3,</sup><sup>4</sup>, <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Turcatti%20G%22%5BAuthor%5D" class="usa-link" aria-describedby="id7"><span class="name western">Gerardo Turcatti</span></a><div hidden="hidden" id="id7"> <h3><span class="name western">Gerardo Turcatti</span></h3> <div class="p"> <sup>2</sup>Biomolecular Screening Facility, School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL), Station 15, Lausanne, 1015 Switzerland </div> <div class="p">Find articles by <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Turcatti%20G%22%5BAuthor%5D" class="usa-link"><span class="name western">Gerardo Turcatti</span></a> </div> </div> <sup>2</sup>, <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Greber%20UF%22%5BAuthor%5D" class="usa-link" aria-describedby="id8"><span class="name western">Urs F Greber</span></a><div hidden="hidden" id="id8"> <h3><span class="name western">Urs F Greber</span></h3> <div class="p"> <sup>1</sup>Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland </div> <div class="p">Find articles by <a href="https://pubmed.ncbi.nlm.nih.gov/?term=%22Greber%20UF%22%5BAuthor%5D" class="usa-link"><span class="name western">Urs F Greber</span></a> </div> </div> <sup>1,</sup><sup>✉</sup> </div> <ul class="d-buttons inline-list"> <li><button class="d-button" aria-controls="aip_a" aria-expanded="false">Author information</button></li> <li><button class="d-button" aria-controls="anp_a" aria-expanded="false">Article notes</button></li> <li><button class="d-button" aria-controls="clp_a" aria-expanded="false">Copyright and License information</button></li> </ul> <div class="d-panels font-secondary-light"> <div id="aip_a" class="d-panel p" style="display: none"> <div class="p" id="Aff1"> <sup>1</sup>Department of Molecular Life Sciences, University of Zurich (UZH), Winterthurerstrasse, 190, 8057 Zurich Switzerland </div> <div id="Aff2"> <sup>2</sup>Biomolecular Screening Facility, School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL), Station 15, Lausanne, 1015 Switzerland </div> <div id="Aff3"> <sup>3</sup>MRC Laboratory for Molecular Cell Biology, University College London, Gower St, London, WC1E 6BT United Kingdom </div> <div id="Aff4"> <sup>4</sup>Artificial Intelligence for Life Sciences CIC, 40 Gowers walk, London, E1 8BH United Kingdom </div> <div class="author-notes p"><div class="fn" id="_fncrsp93pmc__"> <sup>✉</sup><p class="display-inline">Corresponding author.</p> </div></div> </div> <div id="anp_a" class="d-panel p" style="display: none"><div class="notes p"><section id="historyarticle-meta1" class="history"><p>Received 2020 May 18; Accepted 2020 Jul 16; Collection date 2020.</p></section></div></div> <div id="clp_a" class="d-panel p" style="display: none"> <div>© The Author(s) 2020</div> <p><strong>Open Access</strong> This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit <a href="http://creativecommons.org/licenses/by/4.0/" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">http://creativecommons.org/licenses/by/4.0/</a>.</p> <p>The Creative Commons Public Domain Dedication waiver <a href="http://creativecommons.org/publicdomain/zero/1.0/" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">http://creativecommons.org/publicdomain/zero/1.0/</a> applies to the metadata files associated with this article.</p> <div class="p"><a href="/about/copyright/" class="usa-link">PMC Copyright notice</a></div> </div> </div> <div>PMCID: PMC7423605 PMID: <a href="https://pubmed.ncbi.nlm.nih.gov/32788590/" class="usa-link">32788590</a> </div> </div></section></section><section aria-label="Article content"><section class="body main-article-body"><section class="abstract" id="Abs1"><h2>Abstract</h2> <p id="Par1">Human adenoviruses (HAdVs) are fatal to immuno-suppressed individuals, but no effective anti-HAdV therapy is available. Here, we present a novel image-based high-throughput screening (HTS) platform, which scores the full viral replication cycle from virus entry to dissemination of progeny and second-round infections. We analysed 1,280 small molecular weight compounds of the Prestwick Chemical Library (PCL) for interference with HAdV-C2 infection in a quadruplicate, blinded format, and performed robust image analyses and hit filtering. We present the entire set of the screening data including all images, image analyses and data processing pipelines. The data are made available at the Image Data Resource (IDR, idr0081). Our screen identified Nelfinavir mesylate as an inhibitor of HAdV-C2 multi-round plaque formation, but not single round infection. Nelfinavir has been FDA-approved for anti-retroviral therapy in humans. Our results underscore the power of image-based full cycle infection assays in identifying viral inhibitors with clinical potential.</p> <section id="kwd-group1" class="kwd-group"><p><strong>Subject terms:</strong> Adenovirus, Drug development, High-throughput screening</p></section></section><section class="abstract" id="Abs2"><hr class="headless"> <section class="tw xbox font-sm" id="Taba"><div class="tbl-box p" tabindex="0"><table class="content" frame="hsides" rules="groups"><tbody> <tr> <td colspan="1" rowspan="1">Measurement(s)</td> <td colspan="1" rowspan="1">nucleus • number of infected nuclei • infection index (number of infected nuclei/number of nuclei) • Plaque • fluorescent reporter intensity • percent cell viability</td> </tr> <tr> <td colspan="1" rowspan="1">Technology Type(s)</td> <td colspan="1" rowspan="1">epifluorescence microscopy • compound toxicity assay</td> </tr> <tr> <td colspan="1" rowspan="1">Factor Type(s)</td> <td colspan="1" rowspan="1">compound treatment</td> </tr> <tr> <td colspan="1" rowspan="1">Sample Characteristic - Organism</td> <td colspan="1" rowspan="1">Human mastadenovirus C</td> </tr> </tbody></table></div> <div class="p text-right font-secondary"><a href="table/Taba/" class="usa-link" target="_blank" rel="noopener noreferrer">Open in a new tab</a></div></section><p id="Par3">Machine-accessible metadata file describing the reported data: 10.6084/m9.figshare.12594470</p></section><section id="Sec1"><h2 class="pmc_sec_title">Background & Summary</h2> <p id="Par4">Human adenoviruses (HAdVs) affect the respiratory, urinary and gastrointestinal tracts and the eyes. They cause morbidity and mortality, especially to immuno-compromised patients<sup><a href="#CR1" class="usa-link" aria-describedby="CR1">1</a>,<a href="#CR2" class="usa-link" aria-describedby="CR2">2</a></sup> as indicated by a recent outbreak in the USA killing 12 children, or a recent case of meningoencephalitis in a middle-aged woman in the USA<sup><a href="#CR3" class="usa-link" aria-describedby="CR3">3</a></sup>. HAdVs have a high prevalence<sup><a href="#CR4" class="usa-link" aria-describedby="CR4">4</a>–<a href="#CR7" class="usa-link" aria-describedby="CR7">7</a></sup> and are broadly used as gene therapy and vaccination vectors as well as oncolytic viruses<sup><a href="#CR8" class="usa-link" aria-describedby="CR8">8</a>–<a href="#CR10" class="usa-link" aria-describedby="CR10">10</a></sup>. The high seroprevalence of HAdV-C2 and C5 (species C, types 2 and 5) underlines that HAdV infections are asymptomatic in healthy individuals, but persist in mucosal lymphocytes, and thereby pose a risk for immunosuppressed patients undergoing stem cell transplantation<sup><a href="#CR11" class="usa-link" aria-describedby="CR11">11</a>,<a href="#CR12" class="usa-link" aria-describedby="CR12">12</a></sup>. More than 100 HAdV genotypes are grouped into seven species based on hemagglutination assays and genome sequences<sup><a href="#CR13" class="usa-link" aria-describedby="CR13">13</a>,<a href="#CR14" class="usa-link" aria-describedby="CR14">14</a></sup>. Types of the species A, F and G replicate in the gastrointestinal tract, B, C and E in the respiratory tract, and B and D in the conjunctiva of the eyes. Notably, species B members have a broad tropism, including kidney and hematopoietic cells<sup><a href="#CR6" class="usa-link" aria-describedby="CR6">6</a>,<a href="#CR12" class="usa-link" aria-describedby="CR12">12</a></sup>.</p> <p id="Par5">HAdV has a double-stranded DNA genome of ~36 kbp tightly packaged into an icosahedral protein capsid of about 90 nm in diameter<sup><a href="#CR15" class="usa-link" aria-describedby="CR15">15</a>,<a href="#CR16" class="usa-link" aria-describedby="CR16">16</a></sup>. HAdV-C2 and C5 enter cells by receptor-mediated endocytosis, shed minor capsid proteins, expose the membrane lytic protein, penetrate the endosomal membrane and are transported to the nuclear membrane, where they uncoat and release their genome to the nucleus<sup><a href="#CR17" class="usa-link" aria-describedby="CR17">17</a>–<a href="#CR20" class="usa-link" aria-describedby="CR20">20</a></sup>. In the nucleus, the viral genome gives rise to the immediate early viral mRNA encoding the E1A protein, which transactivates the subviral promoters, drives lytic infection and maintains genome persistence in presence of interferon<sup><a href="#CR21" class="usa-link" aria-describedby="CR21">21</a>–<a href="#CR23" class="usa-link" aria-describedby="CR23">23</a></sup>. Proteolytically matured HAdV progeny is released upon rupture of the nuclear envelope and the plasma membrane<sup><a href="#CR24" class="usa-link" aria-describedby="CR24">24</a>–<a href="#CR26" class="usa-link" aria-describedby="CR26">26</a></sup>.</p> <p id="Par6">Currently, there is no effective therapy available against HAdV disease. The standard of care is the nucleoside analogue Cidofovir, with poor clinical efficacy<sup><a href="#CR6" class="usa-link" aria-describedby="CR6">6</a>,<a href="#CR27" class="usa-link" aria-describedby="CR27">27</a></sup>. The problem is exacerbated by the shortage of a suitable small animal model for HAdV disease, although Syrian Hamsters are susceptible to HAdV-C infection and give rise to viral progeny<sup><a href="#CR28" class="usa-link" aria-describedby="CR28">28</a></sup>. Here, we developed an image-based procedure to identify novel inhibitors of HAdV infection in cell culture. We used the commercially available Prestwick Chemical Library (PCL) comprising 1,280 off-patent mostly FDA-approved small molecules (listed in PCL compounds tested in the screening procedure<sup><a href="#CR29" class="usa-link" aria-describedby="CR29">29</a></sup>). The PCL comprises compounds against diseases including infection and cancer<sup><a href="#CR30" class="usa-link" aria-describedby="CR30">30</a>,<a href="#CR31" class="usa-link" aria-describedby="CR31">31</a></sup>, collected at <a href="http://prestwickchemical.com/libraries-publications" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">prestwickchemical.com/libraries-publications</a>.</p> <p id="Par7">We performed a phenotypic screen against HAdV-C2 infection employing automated fluorescence microscopy and image-based scoring of the progression of multi-round infections using the Plaque2.0 software<sup><a href="#CR32" class="usa-link" aria-describedby="CR32">32</a></sup> (Fig. <a href="#Fig1" class="usa-link">1a,b</a>). For representative images in 384 well plates, see Fig. <a href="#Fig1" class="usa-link">1c</a>. The procedure features robust imaging, image analysis and data processing, as concluded from two parallel workflows carried out at independent institutions, the Department of Molecular Life Sciences at University of Zurich (UZH), and the Biomolecular Screening Facility at Ecole Polytechnique Fédérale de Lausanne (EPFL).</p> <figure class="fig xbox font-sm" id="Fig1"><h3 class="obj_head">Fig. 1.</h3> <p class="img-box line-height-none margin-x-neg-2 tablet:margin-x-0 text-center"><a class="tileshop" target="_blank" href="https://www.ncbi.nlm.nih.gov/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Click%20on%20image%20to%20zoom&p=PMC3&id=7423605_41597_2020_604_Fig1_HTML.jpg"><img class="graphic zoom-in" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d5e/7423605/3243f6712be4/41597_2020_604_Fig1_HTML.jpg" loading="lazy" id="d30e497" height="919" width="642" alt="Fig. 1"></a></p> <div class="p text-right font-secondary"><a href="figure/Fig1/" class="usa-link" target="_blank" rel="noopener noreferrer">Open in a new tab</a></div> <figcaption><p>The compound screening procedure. (<strong>a</strong>) Following assay development, stability and quality testing, the screening of the PCL against HAdV infection was performed. Imaging, image analysis and data processing were independently carried out at UZH and EPFL, before hit ranking. (<strong>b</strong>) Schematic overview of the wet-lab pipeline. PCL compounds and DFT positive control in DMSO as well as DMSO alone as negative control were pre-spotted onto 384-well imaging plates by Echo acoustic liquid handling at 10 nl corresponding to a final concentration of 1.25 µM in 80 µl assay volume/well and stored at −20 °C. Compound-blinded plates are thawed and 4,000 A549 cells/wells seeded. The following day, the cells were inoculated with HAdV-C2-dE3B at 1.77*10<sup>5</sup> genome equivalents/well. Allowing for multiple viral replication rounds, the cells were PFA-fixed at 72 hpi and the nuclei stained with Hoechst 33342. The infection phenotypes were imaged using an epifluorescence HT microscope and scored using Plaque2.0. The data of the four technical replicates were further processed in R or through EPFL-BSF LIMS. (<strong>c</strong>) Exemplary epifluorescence microscopy images of cells in 384-wells stitched to a screening plate overview of 32 replicates of negative (two most left columns) and positive control (two most right columns) and 320 blinded PCL compounds (centre 20 columns). Hoechst-stained nuclei are shown in blue, viral GFP in green. (<strong>d</strong>) Representative 384-well epifluorescence microscopy images of the DMSO negative control (most left), the DFT positive control (most right) and the top hit Nelfinavir mesylate (centre). Empty black triangle indicates a plaque (infection focus) from a productively infected cell. White arrows point out infected cells that did not form a plaque. Hoechst-stained nuclei are shown in blue, infected cells expressing GFP in green. Scale bar is 5 mm.</p></figcaption></figure><p id="Par8">Five phenotypic features scored the effects of the compounds on HAdV-C2-dE3B-GFP-infected human lung cancer epithelial A549 cells – the number of infected and uninfected cell nuclei, the infection index (infected nuclei per total nuclei), the number of plaques (areas of infection foci originating from a single productively infected cell as in non-perturbed infections depicted in Fig. <a href="#Fig1" class="usa-link">1d</a>, left image) and the integrated signal of the infection marker green fluorescence protein (GFP) encoded in the reporter virus genome. All data are available at the Image Data Resource (IDR, idr.openmicroscopy.org), IDR accession number idr0081<sup><a href="#CR33" class="usa-link" aria-describedby="CR33">33</a></sup>, and can be accessed via the IDR web client. Raw and scored infection phenotypes are provided for UZH and EPFL analyses. Rigorous assay development ensured a high data quality, as indicated by mean Z’-factors of 0.52 for the plaque numbers. The screening was performed in four biological replicates at high reproducibility. Compounds that gave significant toxicity in uninfected cells were excluded during hit filtering.</p> <p id="Par9">Imaging, image analysis and scoring by the two independent teams yielded well correlated data and a congruent list of top hits, provided in Table <a href="#Tab1" class="usa-link">1</a>. We identified Nelfinavir mesylate (Nelfinavir in short) as best inhibitor of HAdV infection. As evident in representative images presented in Fig. <a href="#Fig1" class="usa-link">1d</a>, Nelfinavir abolishes the formation of plaques, but not single first round infections. We confirmed the antiviral efficacy of Nelfinavir in a follow-up study<sup><a href="#CR34" class="usa-link" aria-describedby="CR34">34</a></sup>.</p> <section class="tw xbox font-sm" id="Tab1"><h3 class="obj_head">Table 1.</h3> <div class="caption p"><p>Summary of screening controls and top hits.</p></div> <div class="tbl-box p" tabindex="0"><table class="content" frame="hsides" rules="groups"> <thead> <tr> <th rowspan="3" colspan="1">Barcode</th> <th rowspan="3" colspan="1">Plate</th> <th colspan="5" rowspan="1">UZH</th> </tr> <tr><th colspan="5" rowspan="1">3 sigma</th></tr> <tr> <th colspan="1" rowspan="1">number Of Nuclei</th> <th colspan="1" rowspan="1">number Of Infected Nuclei</th> <th colspan="1" rowspan="1">infection Index</th> <th colspan="1" rowspan="1">number Of Plaques</th> <th colspan="1" rowspan="1">total Virus Intensity</th> </tr> <tr> <th rowspan="3" colspan="1">Barcode</th> <th rowspan="3" colspan="1">Plate</th> <th colspan="5" rowspan="1">UZH</th> </tr> <tr><th colspan="5" rowspan="1">2 sigma</th></tr> <tr> <th colspan="1" rowspan="1">number Of Nuclei</th> <th colspan="1" rowspan="1">numberOf Infected Nuclei</th> <th colspan="1" rowspan="1">infection Index</th> <th colspan="1" rowspan="1">numberOf Plaques</th> <th colspan="1" rowspan="1">total Virus Intensity</th> </tr> <tr> <th rowspan="3" colspan="1">Barcode</th> <th rowspan="3" colspan="1">Plate</th> <th colspan="5" rowspan="1">EPFL</th> </tr> <tr><th colspan="5" rowspan="1">3 sigma</th></tr> <tr> <th colspan="1" rowspan="1">number Of Nuclei</th> <th colspan="1" rowspan="1">number Of Infected Nuclei</th> <th colspan="1" rowspan="1">infection Index</th> <th colspan="1" rowspan="1">number Of Plaques</th> <th colspan="1" rowspan="1">total Virus Intensity</th> </tr> <tr> <th rowspan="3" colspan="1">Barcode</th> <th rowspan="3" colspan="1">Plate</th> <th colspan="5" rowspan="1">EPFL</th> </tr> <tr><th colspan="5" rowspan="1">2 sigma</th></tr> <tr> <th colspan="1" rowspan="1">number Of Nuclei</th> <th colspan="1" rowspan="1">number Of Infected Nuclei</th> <th colspan="1" rowspan="1">infection Index</th> <th colspan="1" rowspan="1">number Of Plaques</th> <th colspan="1" rowspan="1">total Virus Intensity</th> </tr> </thead> <tbody> <tr> <td colspan="1" rowspan="1">BSF018104</td> <td colspan="1" rowspan="1">Za</td> <td colspan="1" rowspan="1">−1.11</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">0.57</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">0.07</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018105</td> <td colspan="1" rowspan="1">Zb</td> <td colspan="1" rowspan="1">−8.10</td> <td colspan="1" rowspan="1">0.30</td> <td colspan="1" rowspan="1">0.45</td> <td colspan="1" rowspan="1">0.39</td> <td colspan="1" rowspan="1">-0.07</td> </tr> <tr> <td colspan="1" rowspan="1">Mean</td> <td colspan="1" rowspan="1"></td> <td colspan="1" rowspan="1">−4.61</td> <td colspan="1" rowspan="1">0.40</td> <td colspan="1" rowspan="1">0.51</td> <td colspan="1" rowspan="1">0.44</td> <td colspan="1" rowspan="1">0.00</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018104</td> <td colspan="1" rowspan="1">Za</td> <td colspan="1" rowspan="1">−0.41</td> <td colspan="1" rowspan="1">0.67</td> <td colspan="1" rowspan="1">0.71</td> <td colspan="1" rowspan="1">0.67</td> <td colspan="1" rowspan="1">0.07</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018105</td> <td colspan="1" rowspan="1">Zb</td> <td colspan="1" rowspan="1">−5.07</td> <td colspan="1" rowspan="1">0.53</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.59</td> <td colspan="1" rowspan="1">-0.07</td> </tr> <tr> <td colspan="1" rowspan="1">Mean</td> <td colspan="1" rowspan="1"></td> <td colspan="1" rowspan="1">−2.74</td> <td colspan="1" rowspan="1">0.60</td> <td colspan="1" rowspan="1">0.67</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.00</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018104</td> <td colspan="1" rowspan="1">Za</td> <td colspan="1" rowspan="1">−1.20</td> <td colspan="1" rowspan="1">0.36</td> <td colspan="1" rowspan="1">0.47</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.08</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018105</td> <td colspan="1" rowspan="1">Zb</td> <td colspan="1" rowspan="1">−1.23</td> <td colspan="1" rowspan="1">0.27</td> <td colspan="1" rowspan="1">0.32</td> <td colspan="1" rowspan="1">0.44</td> <td colspan="1" rowspan="1">-0.04</td> </tr> <tr> <td colspan="1" rowspan="1">Mean</td> <td colspan="1" rowspan="1"></td> <td colspan="1" rowspan="1">−1.22</td> <td colspan="1" rowspan="1">0.32</td> <td colspan="1" rowspan="1">0.40</td> <td colspan="1" rowspan="1">0.48</td> <td colspan="1" rowspan="1">0.02</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018104</td> <td colspan="1" rowspan="1">Za</td> <td colspan="1" rowspan="1">−0.47</td> <td colspan="1" rowspan="1">0.58</td> <td colspan="1" rowspan="1">0.64</td> <td colspan="1" rowspan="1">0.68</td> <td colspan="1" rowspan="1">0.38</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018105</td> <td colspan="1" rowspan="1">Zb</td> <td colspan="1" rowspan="1">−0.49</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.55</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.31</td> </tr> <tr> <td colspan="1" rowspan="1">Mean</td> <td colspan="1" rowspan="1"></td> <td colspan="1" rowspan="1">−0.48</td> <td colspan="1" rowspan="1">0.55</td> <td colspan="1" rowspan="1">0.60</td> <td colspan="1" rowspan="1">0.66</td> <td colspan="1" rowspan="1">0.35</td> </tr> </tbody> </table></div> <div class="p text-right font-secondary"><a href="table/Tab1/" class="usa-link" target="_blank" rel="noopener noreferrer">Open in a new tab</a></div> <div class="tw-foot p"><div class="fn" id="_fn_p12"><p>Mean corresponds to the means over four biological replicates of PCL compound and 16 biological replicates each carrying 32 technical replicates for each control. Neg. ctr. refers to solvent control (DMSO), pos. ctr. to DFT-treated wells. Normalized indicates the mean read-outs of each compound relative to the mean of the positive control over all replicates. Toxicity was accessed by PrestoBlue assay of 3.5-day treatment of uninfected A549 cells as well as by the nuclei Z’-factor in the screen. Hits were selected for low toxicity and high inhibitory effects compared to solvent control samples. Note that compounds were scored toxic, if they showed significant toxicity in either of the assays.</p></div></div></section></section><section id="Sec2"><h2 class="pmc_sec_title">Methods</h2> <section id="Sec3"><h3 class="pmc_sec_title">Virus</h3> <p id="Par10">HAdV-C2-dE3B-GFP was produced as described<sup><a href="#CR24" class="usa-link" aria-describedby="CR24">24</a></sup> and fully sequenced<sup><a href="#CR35" class="usa-link" aria-describedby="CR35">35</a></sup>. In brief, the virus was generated by exchange of the viral E3B genome region with a reporter cassette harbouring the enhanced green fluorescent protein (GFP) under the immediate early Cytomegalovirus (CMV) promoter<sup><a href="#CR24" class="usa-link" aria-describedby="CR24">24</a></sup>. The virus was grown in A549 cells and purified by double caesium chloride gradient centrifugation<sup><a href="#CR36" class="usa-link" aria-describedby="CR36">36</a></sup>. Aliquots supplemented with 10% glycerol (v/v) were stored at −80 °C. HAdV-C2-dE3B-GFP was found to be homogeneous by SDS-PAGE and negative-stain analyses by transmission electron microscopy.</p></section><section id="Sec4"><h3 class="pmc_sec_title">Cell culture</h3> <p id="Par11">A549 (human adenocarcinomic alveolar basal epithelium) cells were obtained from the American Type Culture Collection (ATCC), Manassas, USA. The cells were maintained in full medium: high glucose Dulbecco Modified Eagle Medium (DMEM; Thermo Fisher Scientific, Waltham, USA) containing 7.5% fetal bovine serum (FBS, Invitrogen, Carlsbad, USA), 1% L-glutamine (Sigma-Aldrich, St. Louis, USA) and 1% penicillin streptomycin (Sigma-Aldrich, St. Louis, USA) and subcultured following PBS washing and trypsinisation (Trypsin-EDTA, Sigma-Aldrich, St. Louis, USA) weekly. Cells were grown at standard conditions (37 °C, 5% CO<sub>2</sub>, 95% humidity) and passage number kept below 20.</p></section><section id="Sec5"><h3 class="pmc_sec_title">Preparation of pre-plates</h3> <p id="Par12">Ten µl 0.0125% DMSO in PBS was spotted on all 384 wells each of imaging-compatible 384-well plates (Matrix plates #4332, Thermo Fisher Scientific, Waltham, USA) using a Matrix WellMate dispenser and normal bore Matrix WellMate tubing cartridges (Thermo Fisher Scientific, Waltham, USA). Plates were sealed and stored at −20 °C.</p></section><section id="Sec6"><h3 class="pmc_sec_title">Blinding</h3> <p id="Par13">The PCL compound arrangement as dispensed by EPFL in four subset plates <em>A</em> - <em>D</em> comprising each screening set replicate <em>1</em>–4 was blinded and replaced by UZH with internal identifier (Raw Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH and Processed Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH<sup><a href="#CR29" class="usa-link" aria-describedby="CR29">29</a></sup>, <em>compoundIdentifier</em> 1 to 1280). The identity of the compounds was only disclosed after the screening process and hit filtering (Raw Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH and Processed Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH<sup><a href="#CR29" class="usa-link" aria-describedby="CR29">29</a></sup> and Table <a href="#Tab1" class="usa-link">1</a>, <em>PCL_ID</em> Prestw-1 to Prestw-1804 and <em>compoundName</em>).</p></section><section id="Sec7"><h3 class="pmc_sec_title">Compounds</h3> <p id="Par14">The PCL was obtained from Prestwick Chemical (Illkirch, France). 3′-deoxy-3′-fluorothymidine (DFT, CAS number 25526-93-6) was obtained from Toronto Research Chemical, North York, Canada. All compounds were dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich, St. Louis, USA) at a final stock concentration of 10 mM and stored at −20 °C.</p></section><section id="Sec8"><h3 class="pmc_sec_title">PrestoBlue toxicity assay</h3> <p id="Par15">Toxicity of the PCL compounds on uninfected A549 cells was assessed using the PrestoBlue Cell Viability reagent (Thermo Fisher Scientific, Waltham, USA). PrestoBlue<sup><a href="#CR37" class="usa-link" aria-describedby="CR37">37</a>,<a href="#CR38" class="usa-link" aria-describedby="CR38">38</a></sup> is a cell viability and cytotoxicity indicator based on resazurin. Resazurin is reduced to resorufin in cellular respiration by accepting electrons from NAPDH, FADH, FMNH, NADH and cytochromes. This reduction causes PrestoBlue to change from a non-fluorescent to a strongly fluorescent form. The conversion of PrestoBlue is proportional to the number of metabolically active cells and can be evaluated quantitatively using fluorescence or absorbance measurements. Briefly, following 3.5-day continuous treatment of A549 cells with compounds at concentrations and cell densities as in the screening protocol, 10% final PrestoBlue was added to each well and incubated for 1 h at standard cell incubation conditions. Fluorescence intensity (bottom-read) was measured using a multi-well plate reader (Tecan Infinite F500, Tecan, Männedorf, Switzerland) with excitation at 560/10 nm, emission at 590/10 nm at a fixed gain. Doxorubicin hydrochloride (Prestw-438, Prestwick Chemical, Illkirch, France) was used as a positive control for cytotoxicity, at a final concentration of 10 µM, and the corresponding concentration of the drug solvent DMSO was used as a negative control. The full PCL library was tested on duplicated plates. The EPFL-BSF in-house Laboratory Information Management System (LIMS) was used for data processing and statistical validation. First, raw PrestoBlue readings were normalized per plate to negative control values at 0 and positive controls at 1. Then, the normalized values of both duplicates were averaged. Assay quality was assessed for each plate through the Z’-factor calculation. Compounds were considered toxic, when the normalized value for all replicates was higher than the average + 3σ (standard deviation, SD) of the DMSO negative control for the corresponding plate. Scores and score SD were then calculated for hit compounds by averaging normalized value for all replicates.</p></section><section id="Sec9"><h3 class="pmc_sec_title">Preparation of plates for Z’-factor and drug screening</h3> <p id="Par16">Ten nl of 10 mM PCL compounds, the nucleoside analogue DFT positive control (all dissolved in DMSO) and DMSO only as negative control were pre-spotted on imaging-compatible 384-well plates (Falcon plates, Corning Inc., New York, USA) using an Echo acoustic liquid handling system (Labcyte, San Jose, USA) by the EPFL-BSF, sealed and stored at −20 °C. Each Z’-factor 384-well plate consisted of 192 technical replicates of positive and negative controls, each. Each screening plate set consisted of four subset plates <em>A</em> to <em>D</em>. Each screening plate comprised 32 technical replicates of positive and negative controls, each, and 320 PCL compounds.</p></section><section id="Sec10"><h3 class="pmc_sec_title">Wet-lab screening pipeline</h3> <p id="Par17">The screening was performed in four independent biological replicates 1–4. Liquid handling was performed using a Matrix WellMate dispenser and Matrix WellMate tubing cartridges (Thermo Fisher Scientific, Waltham, USA). Prior to usage, tubings were rinsed with 125 ml autoclaved double-distilled (dd) H<sub>2</sub>O followed by 125 ml autoclaved PBS. Pre-spotted compound plates were thawed at room temperature (RT) for 30 min, briefly centrifuged before compounds were dissolved in 10 µl/well of PBS. 4,000 A549 cells/well in 60 µl full medium were seeded onto the compounds using standard bore tubing cartridges. Following cell adhesion over night, the cells were inoculated with 1.77*10<sup>5</sup> genome equivalents per well of HAdV-C2-dE3B-GFP in 10 µl of full medium using bovine serum albumin (BSA, cell-culture grade, Sigma-Aldrich, St. Louis, USA)-blocked small bore tubing cartridges. The final compound concentration was 1.25 µM at 0.0125% DMSO. Infection was allowed to progress over multiple infection rounds for 72 h giving rise to foci of infected cells originating from a single first round infected cell. Cells were fixed for 1 h at RT by addition of 26.6 µL 16% PFA and 4 µg/ml Hoechst 33342 (Sigma-Aldrich, St. Louis, USA) in PBS using standard bore tubing cartridges. Cells were washed three times with PBS before PBS supplemented with 0.02% N<sub>3</sub> was added and plates were sealed for long-term storage at 4 °C. Following usage, tubings were rinsed with 125 ml autoclaved ddH<sub>2</sub>O followed by 125 ml autoclaved PBS and autoclaved for re-usage.</p></section><section id="Sec11"><h3 class="pmc_sec_title">Imaging</h3> <p id="Par18">Nuclei stained with Hoechst 33342 (DAPI channel) and viral GFP (FITC channel) were imaged on two devices. At UZH, plates were imaged on an IXM-C automated high-throughput fluorescence microscope (Molecular Devices, San Jose, USA) using MetaXpress (version 6.2, Molecular Devices, San Jose, USA) and a 4x air objective (Nikon S Fluor, 0.20 NA, 15.5 mm WD, Nikon Instruments, Minato, Japan) at widefield mode. Images of 2,048<sup>2</sup> px at 1.72 µm/px resolution were acquired on an Andor sCMOS camera (Oxford Instruments, Abingdon, UK). Exposure times: DAPI 150 ms, FITC 20 ms. At EPFL, images were acquired on a IN Cell 2200 automated high-throughput fluorescence microscope (GE Healthcare, Chicago, USA) using IN Cell Analyzer (version 6.2, GE Healthcare, Chicago, USA) and a 4x air objective (Nikon Plan Apo, 0.20 NA, 15.7 mm WD, Nikon Instruments, Minato, Japan) at widefield mode. Image size 2,048<sup>2</sup> px at 1.625 µm/px resolution acquired on an Andor sCMOS camera. Exposure times: DAPI 300 ms, FITC 40 ms.</p></section><section id="Sec12"><h3 class="pmc_sec_title">Image analysis</h3> <p id="Par19">The infection phenotype for each well was quantified by Plaque2.0<sup><a href="#CR32" class="usa-link" aria-describedby="CR32">32</a></sup> (<a href="https://github.com/plaque2/matlab/tree/antivir" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">https://github.com/plaque2/matlab/tree/antivir</a>) via five main read-outs: number of nuclei, number of infected nuclei, the ratio between infected and total nuclei referred to as infection index, number of multi-round infection foci termed plaques (plaque forming unit(s), pfu) and the integrated viral transgenic GFP intensity. Plaque2.0 parameters were optimized independently at UZH and EPFL for the data acquired at the respective institution. Well- and object-based read-outs are provided in the Plaque2.0 output files.</p></section><section id="Sec13"><h3 class="pmc_sec_title">Z’-factor calculation</h3> <p id="Par20">The Z’-factor was computed using R version 3.3.2<sup><a href="#CR39" class="usa-link" aria-describedby="CR39">39</a></sup> according to Eq. (<a href="#Equ1" class="usa-link">1</a>)</p> <table class="disp-formula p" id="Equ1"><tr> <td class="formula"><math id="M2" display="block"><msup><mrow><mi>Z</mi></mrow><mrow><mo>′</mo></mrow></msup><mo>=</mo><mn>1</mn><mo>−</mo><mfrac><mrow><mrow><mo>(</mo><mrow><mn>3</mn><msub><mrow><mi>σ</mi></mrow><mrow><mo>+</mo></mrow></msub><mo>+</mo><mn>3</mn><msub><mrow><mi>σ</mi></mrow><mrow><mo>−</mo></mrow></msub></mrow><mo>)</mo></mrow></mrow><mrow><mfenced close="∣" open="∣"><mrow><msub><mrow><mi>μ</mi></mrow><mrow><mo>+</mo></mrow></msub><mo>−</mo><msub><mrow><mi>μ</mi></mrow><mrow><mo>−</mo></mrow></msub></mrow></mfenced></mrow></mfrac></math></td> <td class="label">1</td> </tr></table> <p>where σ<sub>+</sub> is the SD of the positive control, σ<sub>-</sub> is the SD of the negative control, μ<sub>+</sub> the mean of the positive control and μ<sub>-</sub> the mean of the negative control.</p></section><section id="Sec14"><h3 class="pmc_sec_title">Screening data processing</h3> <p id="Par21">Plaque2.0 results were further processed and filtered. At UZH, results were processed in R version 3.3.2<sup><a href="#CR39" class="usa-link" aria-describedby="CR39">39</a></sup>, EPFL used KNIME version 3.4.0<sup><a href="#CR40" class="usa-link" aria-describedby="CR40">40</a></sup> as well as the EPFL-BSF in-house LIMS. Mean infection scores over the five Plaque2.0 read-outs of the four biological replicates of each PCL compound and the 16 biological replicates containing 32 technical replicates of positive and negative control, each, were calculated. Each compound’s scores were normalized by the mean score of the DMSO negative control of the respective plate. Only non-toxic, effective PCL compounds were considered as HAdV inhibitor candidates. Non-toxic compounds were filtered by applying an inclusive μ<sub>+</sub> (mean of the negative control) ± 2σ (SD of the negative control) threshold for number of nuclei. Efficacy was filtered by applying an excluding μ<sub>+</sub> ± 3σ threshold for the infection scores (number of infected nuclei, infection index, number of plaques or integrated GFP intensity). Subsequently, compounds exhibiting significant toxicity to noninfected cells were excluded.</p></section></section><section id="Sec15"><h2 class="pmc_sec_title">Data Records</h2> <section id="Sec16"><h3 class="pmc_sec_title">Data structure and repositories</h3> <p id="Par22">The screening data comprise the information collected during assay development, including stability, quality and screening of the PCL itself. The latter two were imaged on two different microscopes. We provide the parameters used for Plaque2.0 image analysis, and the code for the subsequent hit filtering in R. The data available for download at the IDR, accession number idr0081<sup><a href="#CR33" class="usa-link" aria-describedby="CR33">33</a></sup>, are structured as outlined in Fig. <a href="#Fig2" class="usa-link">2a</a>. For download instructions, see idr.openmicroscopy.org/about/download. Moreover, the data can be viewed conveniently on the IDR web client (idr.openmicroscopy.org/webclient), where it is structured as depicted in Fig. <a href="#Fig2" class="usa-link">2b</a>. Additionally, an annotated list of the PCL compounds as well as raw and scored screening data are available on figshare<sup><a href="#CR29" class="usa-link" aria-describedby="CR29">29</a></sup> as.txt files.</p> <figure class="fig xbox font-sm" id="Fig2"><h4 class="obj_head">Fig. 2.</h4> <p class="img-box line-height-none margin-x-neg-2 tablet:margin-x-0 text-center"><a class="tileshop" target="_blank" href="https://www.ncbi.nlm.nih.gov/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Click%20on%20image%20to%20zoom&p=PMC3&id=7423605_41597_2020_604_Fig2_HTML.jpg"><img class="graphic zoom-in" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d5e/7423605/81851ed9d681/41597_2020_604_Fig2_HTML.jpg" loading="lazy" id="d30e1239" height="458" width="660" alt="Fig. 2"></a></p> <div class="p text-right font-secondary"><a href="figure/Fig2/" class="usa-link" target="_blank" rel="noopener noreferrer">Open in a new tab</a></div> <figcaption><p>Project data structure available at IDR, accession number idr0081<sup><a href="#CR33" class="usa-link" aria-describedby="CR33">33</a></sup>. (<strong>a</strong>) In the data provided for download, there are three sub-folders for <em>1-prePlates</em>, <em>2-ZPlates</em> and <em>3-Screen</em>. The latter two contain both the images and analyses generated by UZH and EPFL. (<strong>b</strong>) The data provided for viewing are divided into five screens: <em>screenA</em> contains the pre-plates and <em>screenB</em> and <em>screenC</em> consist of the Z’-factor plates imaged and analysed at UZH and EPFL, respectively. <em>screenD</em> and <em>screenE</em> provide the screening data obtained at UZH and EPFL, respectively.</p></figcaption></figure></section><section id="Sec17"><h3 class="pmc_sec_title">Data sets and file types</h3> <p id="Par23">The data provided for download consists of three data sets <em>1</em> to <em>3</em> (see Fig. <a href="#Fig2" class="usa-link">2a</a>).</p> <p id="Par24">- <em>1-prePlates</em> contains layouts (.csv), images (.tif), Plaque2.0 image analysis parameters (.mat) and results (.csv) for the assay stability test plates performed at UZH prior to Z’-factor plates (<em>preZ</em>) and the screen (<em>preScreen</em>).</p> <p id="Par25">- <em>2-ZPlates</em> contains layouts (.csv), images (.tif), Plaque2.0 image analysis parameters (.mat) and results (.csv) for the two Z’-factor plates <em>a</em> and <em>b</em> as imaged and analysed at UZH (<em>Data_UZH</em>) and EPFL (<em>Data_EPFL</em>).</p> <p id="Par26">- <em>3-Screen</em> contains layouts (.csv), images (.tif), Plaque2.0 image analysis parameters (.mat) and results (.csv) for the 16 screening plates (four biological replicates <em>1–4</em>, each consisting of a set of four subset plates <em>A - D</em>) as imaged and analysed at UZH (<em>Data_UZH</em>) and EPFL (<em>Data_EPFL</em>). Moreover, <em>Analysis</em> contains the Plaque2.0 batch processing (<em>AntiVir_batchprocessing.m</em>) and hit filtering pipeline (<em>AntiVir_hitfiltering.R</em>) used by UZH. <em>Analysis</em> also contains the PrestoBlue raw results (.csv) for toxicity in absence of infection.</p> <p id="Par27">The data provided for browsing via the IDR web client are assembled into five screens <em>A</em> to <em>E</em> (see Fig. <a href="#Fig2" class="usa-link">2b</a>).</p> <p id="Par28">- idr0081-study.txt summarizes the overall study and the five screens that were performed.</p> <p id="Par29">- <em>screenA</em> contains the assay stability test plates performed at UZH prior to Z’-factor plates (<em>preZ</em>) and the screen (<em>preScreen</em>). <em>idr0081-screenA-library.txt</em> provides thorough information on the tested compounds including PubChem identifiers and their plate layout. <em>idr0081-screenA-processed.txt</em> presents the results of the Plaque2.0-based image analysis. <em>idr0081-screenA-mean.txt</em> summarises the infection scores per pre plate.</p> <p id="Par30">- s<em>creenB</em> contains the assay quality test plates (Z’-factor plates <em>a</em> and <em>b</em>) performed at UZH. <em>idr0081-screenB-library.txt</em> provides thorough information on the tested compounds including PubChem identifiers and their plate layout. <em>idr0081-screenB-processed.txt</em> presents the results of the Plaque2.0-based image analysis. <em>idr0081-screenB-mean.txt</em> summarises the infection scores per Z’-factor plate.</p> <p id="Par31">- <em>screenC</em> contains the assay quality test plates (Z’-factor plates <em>a</em> and <em>b</em>) performed at EPFL. <em>idr0081-screenC-library.txt</em> provides thorough information on the tested compounds including PubChem identifiers and their plate layout. <em>idr0081-screenC-processed.txt</em> presents the results of the Plaque2.0-based image analysis. <em>idr0081-screenC-mean.txt</em> summarises the infection scores per Z’-factor plate.</p> <p id="Par32">- <em>screenD</em> contains the PCL screening plates (in replicates <em>1</em> to <em>4</em>, consisting of subset plates <em>A</em> to <em>D</em>) performed at UZH. <em>idr0081-screenD-library.txt</em> provides thorough information on the tested compounds including PubChem identifiers and their plate layout. <em>idr0081-screenD-processed.txt</em> presents the results of the Plaque2.0-based image analysis. <em>idr0081-screenB-filtered.txt</em> summarises the infection scores per compound and indicates if it was identified as hit.</p> <p id="Par33">- <em>screenE</em> contains the PCL screening plates (in replicates <em>1</em> to <em>4</em>, consisting of subsets <em>A</em> to <em>D</em>) performed at EPFL. <em>idr0081-screenE-library.txt</em> provides thorough information on the tested compounds including PubChem identifiers and their plate layout. <em>idr0081-screenE-processed.txt</em> presents the results of the Plaque2.0-based image analysis. <em>idr0081-screenE-filtered.txt</em> summarises the infection scores per compound and indicates, if it was identified as hit.</p></section></section><section id="Sec18"><h2 class="pmc_sec_title">Technical Validation</h2> <section id="Sec19"><h3 class="pmc_sec_title">Assay stability</h3> <p id="Par34">The wet-lab screening pipeline was optimized regarding liquid handling, cell seeding, virus inoculum, positive and negative controls, infection time, as well as imaging and image analysis. This ensured a high assay stability and reproducibility. Furthermore, all compounds, especially media and supplements, the BSA for tubing saturation, PFA- and Hoechst-supplemented fixative were prepared as large batch from a single lot and stored as single-use aliquots. Prior to every experiment, assay stability with respect to cell and infection phenotype was tested on pre-plates according to the established wet-lab, imaging and image analysis pipeline. Since the solvent control had already been spotted in 10 µl PBS, no further PBS was added prior to cell seeding. Periodically, the virus stock dilution was tested and adjusted for experiments if necessary.</p></section><section id="Sec20"><h3 class="pmc_sec_title">Assay quality determination: Z’-factor</h3> <p id="Par35">The accuracy of the wet-lab, imaging and image analysis pipeline was assessed by two independently imaged and analysed Z’-factor plates (Table <a href="#Tab2" class="usa-link">2</a> and Fig. <a href="#Fig3" class="usa-link">3</a>). 3σ Z’-factors of <em>numberOfInfectedNuclei</em>, <em>infectionIndex</em> and <em>numberOfPlaques</em> were in the range of 0.30 to 0.57, scoring good to excellent. <em>totalVirusIntensity</em> (Z’-factors between −0.07 to 0.08) were not suitable to identify HAdV infection inhibitors, while <em>numberOfNuclei</em> (Z’-factors between −1.11 to −8.10) was not a useable readout either. Additionally, the Z’-factors were determined for each of the 16 screening plates (Table <a href="#Tab3" class="usa-link">3</a> and Fig. <a href="#Fig4" class="usa-link">4</a>). 3σ Z’-factors of <em>numberOfInfectedNuclei</em>, <em>infectionIndex</em> and <em>numberOfPlaques</em> were in the range of 0.27 to 0.57, scoring good to excellent.</p> <section class="tw xbox font-sm" id="Tab2"><h4 class="obj_head">Table 2.</h4> <div class="caption p"><p>Z’-factor plates.</p></div> <div class="tbl-box p" tabindex="0"><table class="content" frame="hsides" rules="groups"> <thead> <tr> <th rowspan="3" colspan="1">Barcode</th> <th rowspan="3" colspan="1">Plate</th> <th colspan="5" rowspan="1">UZH</th> <th colspan="5" rowspan="1">EPFL</th> </tr> <tr> <th colspan="5" rowspan="1">3 sigma</th> <th colspan="5" rowspan="1">3 sigma</th> </tr> <tr> <th colspan="1" rowspan="1">number Of Nuclei</th> <th colspan="1" rowspan="1">number Of Infected Nuclei</th> <th colspan="1" rowspan="1">infection Index</th> <th colspan="1" rowspan="1">number Of Plaques</th> <th colspan="1" rowspan="1">total Virus Intensity</th> <th colspan="1" rowspan="1">number Of Nuclei</th> <th colspan="1" rowspan="1">number Of Infected Nuclei</th> <th colspan="1" rowspan="1">infection Index</th> <th colspan="1" rowspan="1">number Of Plaques</th> <th colspan="1" rowspan="1">total Virus Intensity</th> </tr> <tr> <th rowspan="3" colspan="1">Barcode</th> <th rowspan="3" colspan="1">Plate</th> <th colspan="5" rowspan="1">UZH</th> <th colspan="5" rowspan="1">EPFL</th> </tr> <tr> <th colspan="5" rowspan="1">2 sigma</th> <th colspan="5" rowspan="1">2 sigma</th> </tr> <tr> <th colspan="1" rowspan="1">number Of Nuclei</th> <th colspan="1" rowspan="1">number Of Infected Nuclei</th> <th colspan="1" rowspan="1">infection Index</th> <th colspan="1" rowspan="1">number Of Plaques</th> <th colspan="1" rowspan="1">total Virus Intensity</th> <th colspan="1" rowspan="1">number Of Nuclei</th> <th colspan="1" rowspan="1">number Of Infected Nuclei</th> <th colspan="1" rowspan="1">infection Index</th> <th colspan="1" rowspan="1">number Of Plaques</th> <th colspan="1" rowspan="1">total Virus Intensity</th> </tr> </thead> <tbody> <tr> <td colspan="1" rowspan="1">BSF018104</td> <td colspan="1" rowspan="1">Za</td> <td colspan="1" rowspan="1">−1.11</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">0.57</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">0.07</td> <td colspan="1" rowspan="1">−1.20</td> <td colspan="1" rowspan="1">0.36</td> <td colspan="1" rowspan="1">0.47</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.08</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018105</td> <td colspan="1" rowspan="1">Zb</td> <td colspan="1" rowspan="1">−8.10</td> <td colspan="1" rowspan="1">0.30</td> <td colspan="1" rowspan="1">0.45</td> <td colspan="1" rowspan="1">0.39</td> <td colspan="1" rowspan="1">−0.07</td> <td colspan="1" rowspan="1">−1.23</td> <td colspan="1" rowspan="1">0.27</td> <td colspan="1" rowspan="1">0.32</td> <td colspan="1" rowspan="1">0.44</td> <td colspan="1" rowspan="1">−0.04</td> </tr> <tr> <td colspan="1" rowspan="1">Mean</td> <td colspan="1" rowspan="1"></td> <td colspan="1" rowspan="1">−4.61</td> <td colspan="1" rowspan="1">0.40</td> <td colspan="1" rowspan="1">0.51</td> <td colspan="1" rowspan="1">0.44</td> <td colspan="1" rowspan="1">0.00</td> <td colspan="1" rowspan="1">−1.22</td> <td colspan="1" rowspan="1">0.32</td> <td colspan="1" rowspan="1">0.40</td> <td colspan="1" rowspan="1">0.48</td> <td colspan="1" rowspan="1">0.02</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018104</td> <td colspan="1" rowspan="1">Za</td> <td colspan="1" rowspan="1">−0.41</td> <td colspan="1" rowspan="1">0.67</td> <td colspan="1" rowspan="1">0.71</td> <td colspan="1" rowspan="1">0.67</td> <td colspan="1" rowspan="1">0.07</td> <td colspan="1" rowspan="1">−0.47</td> <td colspan="1" rowspan="1">0.58</td> <td colspan="1" rowspan="1">0.64</td> <td colspan="1" rowspan="1">0.68</td> <td colspan="1" rowspan="1">0.38</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018105</td> <td colspan="1" rowspan="1">Zb</td> <td colspan="1" rowspan="1">−5.07</td> <td colspan="1" rowspan="1">0.53</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.59</td> <td colspan="1" rowspan="1">−0.07</td> <td colspan="1" rowspan="1">−0.49</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.55</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.31</td> </tr> <tr> <td colspan="1" rowspan="1">Mean</td> <td colspan="1" rowspan="1"></td> <td colspan="1" rowspan="1">−2.74</td> <td colspan="1" rowspan="1">0.60</td> <td colspan="1" rowspan="1">0.67</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.00</td> <td colspan="1" rowspan="1">−0.48</td> <td colspan="1" rowspan="1">0.55</td> <td colspan="1" rowspan="1">0.60</td> <td colspan="1" rowspan="1">0.66</td> <td colspan="1" rowspan="1">0.35</td> </tr> </tbody> </table></div> <div class="p text-right font-secondary"><a href="table/Tab2/" class="usa-link" target="_blank" rel="noopener noreferrer">Open in a new tab</a></div> <div class="tw-foot p"><div class="fn" id="_fn_p41"><p>The quality of the platform was assessed prior to screening of the PCL by two independent Z’-factor plates containing 192 technical replicates of both positive control (1.25 µM DFT) and solvent only control (0.0125% DMSO). Z’-factors for the five Plaque2.0 read-outs<sup><a href="#CR32" class="usa-link" aria-describedby="CR32">32</a></sup> obtained by independent analysis at UZH and EPFL were calculated according to Eq. (<a href="#Equ1" class="usa-link">1</a>) for 3 and 2σ.</p></div></div></section><figure class="fig xbox font-sm" id="Fig3"><h4 class="obj_head">Fig. 3.</h4> <p class="img-box line-height-none margin-x-neg-2 tablet:margin-x-0 text-center"><a class="tileshop" target="_blank" href="https://www.ncbi.nlm.nih.gov/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Click%20on%20image%20to%20zoom&p=PMC3&id=7423605_41597_2020_604_Fig3_HTML.jpg"><img class="graphic zoom-in" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d5e/7423605/36ff0de8bc71/41597_2020_604_Fig3_HTML.jpg" loading="lazy" id="d30e1891" height="722" width="660" alt="Fig. 3"></a></p> <div class="p text-right font-secondary"><a href="figure/Fig3/" class="usa-link" target="_blank" rel="noopener noreferrer">Open in a new tab</a></div> <figcaption><p>Infection score density of positive and negative controls across Z’-factor plates. Distribution of (<strong>a</strong>) <em>numberOfNuclei</em>, (<strong>b</strong>) <em>numberOfInfectedNuclei</em>, (<strong>c</strong>) <em>infectionIndex</em>, (<strong>d</strong>) <em>numberOfPlaques</em> and <strong>e</strong><em>totalVirusIntensity</em> in negative control (0.0125% DMSO) compared to positive control-treated (1.25 µM DFT) samples of the two Z’-factor plates. Dark green and dark grey indicate Z’-factor plate a, light green and grey show Z’-factor plate b. Dashed vertical lines mark mean of 192 technical replicates.</p></figcaption></figure><section class="tw xbox font-sm" id="Tab3"><h4 class="obj_head">Table 3.</h4> <div class="caption p"><p>Z’-factors of screening plates.</p></div> <div class="tbl-box p" tabindex="0"><table class="content" frame="hsides" rules="groups"> <thead> <tr> <th rowspan="3" colspan="1">Barcode</th> <th rowspan="3" colspan="1">Plate</th> <th colspan="5" rowspan="1">UZH</th> <th colspan="5" rowspan="1">EPFL</th> </tr> <tr> <th colspan="5" rowspan="1">3 sigma</th> <th colspan="5" rowspan="1">3 sigma</th> </tr> <tr> <th colspan="1" rowspan="1">number Of Nuclei</th> <th colspan="1" rowspan="1">number Of Infected Nuclei</th> <th colspan="1" rowspan="1">infection Index</th> <th colspan="1" rowspan="1">number Of Plaques</th> <th colspan="1" rowspan="1">totalVirus Intensity</th> <th colspan="1" rowspan="1">number Of Nuclei</th> <th colspan="1" rowspan="1">number Of Infected Nuclei</th> <th colspan="1" rowspan="1">infection Index</th> <th colspan="1" rowspan="1">number Of Plaques</th> <th colspan="1" rowspan="1">total Virus Intensity</th> </tr> </thead> <tbody> <tr> <td colspan="1" rowspan="1">BSF018292</td> <td colspan="1" rowspan="1">1 A</td> <td colspan="1" rowspan="1">−0.13</td> <td colspan="1" rowspan="1">0.58</td> <td colspan="1" rowspan="1">0.58</td> <td colspan="1" rowspan="1">0.59</td> <td colspan="1" rowspan="1">0.35</td> <td colspan="1" rowspan="1">−0.14</td> <td colspan="1" rowspan="1">0.51</td> <td colspan="1" rowspan="1">0.49</td> <td colspan="1" rowspan="1">0.58</td> <td colspan="1" rowspan="1">0.31</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018293</td> <td colspan="1" rowspan="1">1B</td> <td colspan="1" rowspan="1">−0.88</td> <td colspan="1" rowspan="1">0.58</td> <td colspan="1" rowspan="1">0.65</td> <td colspan="1" rowspan="1">0.55</td> <td colspan="1" rowspan="1">0.34</td> <td colspan="1" rowspan="1">−0.35</td> <td colspan="1" rowspan="1">0.51</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.51</td> <td colspan="1" rowspan="1">0.35</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018294</td> <td colspan="1" rowspan="1">1 C</td> <td colspan="1" rowspan="1">−1.01</td> <td colspan="1" rowspan="1">0.62</td> <td colspan="1" rowspan="1">0.62</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.33</td> <td colspan="1" rowspan="1">−0.74</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">0.66</td> <td colspan="1" rowspan="1">0.32</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018295</td> <td colspan="1" rowspan="1">1D</td> <td colspan="1" rowspan="1">−0.34</td> <td colspan="1" rowspan="1">0.56</td> <td colspan="1" rowspan="1">0.54</td> <td colspan="1" rowspan="1">0.45</td> <td colspan="1" rowspan="1">0.16</td> <td colspan="1" rowspan="1">−0.21</td> <td colspan="1" rowspan="1">0.43</td> <td colspan="1" rowspan="1">0.38</td> <td colspan="1" rowspan="1">0.46</td> <td colspan="1" rowspan="1">0.19</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018296</td> <td colspan="1" rowspan="1">2 A</td> <td colspan="1" rowspan="1">−1.35</td> <td colspan="1" rowspan="1">0.64</td> <td colspan="1" rowspan="1">0.67</td> <td colspan="1" rowspan="1">0.55</td> <td colspan="1" rowspan="1">0.30</td> <td colspan="1" rowspan="1">−0.20</td> <td colspan="1" rowspan="1">0.57</td> <td colspan="1" rowspan="1">0.55</td> <td colspan="1" rowspan="1">0.55</td> <td colspan="1" rowspan="1">0.28</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018297</td> <td colspan="1" rowspan="1">2B</td> <td colspan="1" rowspan="1">−3.63</td> <td colspan="1" rowspan="1">0.56</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.45</td> <td colspan="1" rowspan="1">0.14</td> <td colspan="1" rowspan="1">−1.20</td> <td colspan="1" rowspan="1">0.45</td> <td colspan="1" rowspan="1">0.39</td> <td colspan="1" rowspan="1">0.40</td> <td colspan="1" rowspan="1">0.12</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018298</td> <td colspan="1" rowspan="1">2 C</td> <td colspan="1" rowspan="1">−1.81</td> <td colspan="1" rowspan="1">0.60</td> <td colspan="1" rowspan="1">0.58</td> <td colspan="1" rowspan="1">0.49</td> <td colspan="1" rowspan="1">0.24</td> <td colspan="1" rowspan="1">−0.38</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.42</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.19</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018299</td> <td colspan="1" rowspan="1">2D</td> <td colspan="1" rowspan="1">−1.94</td> <td colspan="1" rowspan="1">0.57</td> <td colspan="1" rowspan="1">0.57</td> <td colspan="1" rowspan="1">0.57</td> <td colspan="1" rowspan="1">0.24</td> <td colspan="1" rowspan="1">−0.22</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">0.43</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.20</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018300</td> <td colspan="1" rowspan="1">3 A</td> <td colspan="1" rowspan="1">−1.74</td> <td colspan="1" rowspan="1">0.64</td> <td colspan="1" rowspan="1">0.66</td> <td colspan="1" rowspan="1">0.56</td> <td colspan="1" rowspan="1">0.36</td> <td colspan="1" rowspan="1">−0.54</td> <td colspan="1" rowspan="1">0.55</td> <td colspan="1" rowspan="1">0.51</td> <td colspan="1" rowspan="1">0.59</td> <td colspan="1" rowspan="1">0.34</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018301</td> <td colspan="1" rowspan="1">3B</td> <td colspan="1" rowspan="1">−1.13</td> <td colspan="1" rowspan="1">0.60</td> <td colspan="1" rowspan="1">0.68</td> <td colspan="1" rowspan="1">0.58</td> <td colspan="1" rowspan="1">0.40</td> <td colspan="1" rowspan="1">−0.09</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.57</td> <td colspan="1" rowspan="1">0.59</td> <td colspan="1" rowspan="1">0.40</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018302</td> <td colspan="1" rowspan="1">3 C</td> <td colspan="1" rowspan="1">−4.02</td> <td colspan="1" rowspan="1">0.66</td> <td colspan="1" rowspan="1">0.68</td> <td colspan="1" rowspan="1">0.48</td> <td colspan="1" rowspan="1">0.42</td> <td colspan="1" rowspan="1">−1.07</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.60</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">0.41</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018303</td> <td colspan="1" rowspan="1">3D</td> <td colspan="1" rowspan="1">−2.36</td> <td colspan="1" rowspan="1">0.55</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.51</td> <td colspan="1" rowspan="1">0.36</td> <td colspan="1" rowspan="1">−0.10</td> <td colspan="1" rowspan="1">0.58</td> <td colspan="1" rowspan="1">0.54</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.35</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018304</td> <td colspan="1" rowspan="1">4 A</td> <td colspan="1" rowspan="1">−0.68</td> <td colspan="1" rowspan="1">0.70</td> <td colspan="1" rowspan="1">0.74</td> <td colspan="1" rowspan="1">0.42</td> <td colspan="1" rowspan="1">0.37</td> <td colspan="1" rowspan="1">−0.29</td> <td colspan="1" rowspan="1">0.56</td> <td colspan="1" rowspan="1">0.58</td> <td colspan="1" rowspan="1">0.48</td> <td colspan="1" rowspan="1">0.36</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018305</td> <td colspan="1" rowspan="1">4B</td> <td colspan="1" rowspan="1">−0.17</td> <td colspan="1" rowspan="1">0.71</td> <td colspan="1" rowspan="1">0.74</td> <td colspan="1" rowspan="1">0.51</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">−0.50</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.67</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">0.50</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018306</td> <td colspan="1" rowspan="1">4 C</td> <td colspan="1" rowspan="1">−0.44</td> <td colspan="1" rowspan="1">0.61</td> <td colspan="1" rowspan="1">0.62</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">0.28</td> <td colspan="1" rowspan="1">−0.28</td> <td colspan="1" rowspan="1">0.50</td> <td colspan="1" rowspan="1">0.48</td> <td colspan="1" rowspan="1">0.47</td> <td colspan="1" rowspan="1">0.26</td> </tr> <tr> <td colspan="1" rowspan="1">BSF018307</td> <td colspan="1" rowspan="1">4D</td> <td colspan="1" rowspan="1">−0.77</td> <td colspan="1" rowspan="1">0.63</td> <td colspan="1" rowspan="1">0.70</td> <td colspan="1" rowspan="1">0.42</td> <td colspan="1" rowspan="1">0.41</td> <td colspan="1" rowspan="1">−0.22</td> <td colspan="1" rowspan="1">0.54</td> <td colspan="1" rowspan="1">0.56</td> <td colspan="1" rowspan="1">0.36</td> <td colspan="1" rowspan="1">0.39</td> </tr> <tr> <td colspan="1" rowspan="1">Mean</td> <td colspan="1" rowspan="1"></td> <td colspan="1" rowspan="1">−1.40</td> <td colspan="1" rowspan="1">0.61</td> <td colspan="1" rowspan="1">0.64</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.32</td> <td colspan="1" rowspan="1">−0.41</td> <td colspan="1" rowspan="1">0.53</td> <td colspan="1" rowspan="1">0.51</td> <td colspan="1" rowspan="1">0.52</td> <td colspan="1" rowspan="1">0.31</td> </tr> </tbody> </table></div> <div class="p text-right font-secondary"><a href="table/Tab3/" class="usa-link" target="_blank" rel="noopener noreferrer">Open in a new tab</a></div> <div class="tw-foot p"><div class="fn" id="_fn_p44"><p>The quality of the screening data was assessed for each screening plate based on the 32 technical replicates of both positive control (1.25 µM DFT) and solvent only control (0.0125% DMSO) in each plate. Z’-factors for the five Plaque2.0 read-outs<sup><a href="#CR32" class="usa-link" aria-describedby="CR32">32</a></sup> obtained by independent analysis at UZH and EPFL were calculated according to Eq. (<a href="#Equ1" class="usa-link">1</a>) for 3σ.</p></div></div></section><figure class="fig xbox font-sm" id="Fig4"><h4 class="obj_head">Fig. 4.</h4> <p class="img-box line-height-none margin-x-neg-2 tablet:margin-x-0 text-center"><a class="tileshop" target="_blank" href="https://www.ncbi.nlm.nih.gov/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Click%20on%20image%20to%20zoom&p=PMC3&id=7423605_41597_2020_604_Fig4_HTML.jpg"><img class="graphic zoom-in" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d5e/7423605/090f2dde40ff/41597_2020_604_Fig4_HTML.jpg" loading="lazy" id="d30e2659" height="725" width="658" alt="Fig. 4"></a></p> <div class="p text-right font-secondary"><a href="figure/Fig4/" class="usa-link" target="_blank" rel="noopener noreferrer">Open in a new tab</a></div> <figcaption><p>Infection score density of positive and negative controls across screening replicates. Distribution of (<strong>a</strong>) <em>numberOfNuclei</em>, (<strong>b</strong>) <em>numberOfInfectedNuclei</em>, (<strong>c</strong>) <em>infectionIndex</em>, (<strong>d</strong>) <em>numberOfPlaques</em> and <strong>e</strong><em>totalVirusIntensity</em> in negative control (0.0125% DMSO in grey) compared to positive control-treated (1.25 µM DFT in green) samples of the screening sets. Replicates 1 to 4 indicated by colour shading are each comprised of four set plates containing 32 technical replicates per control. The dashed vertical lines indicate the corresponding mean values.</p></figcaption></figure></section><section id="Sec21"><h3 class="pmc_sec_title">Independent analysis and filtering</h3> <p id="Par36">Imaging, image analysis and screening data processing were performed by two independent research teams at UZH and EPFL, as depicted in Fig. <a href="#Fig1" class="usa-link">1</a>. Raw and scored infection phenotypes are shown for UZH and EPFL analyses (Raw Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH, Processed Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH and Raw Plaque-2.0 infection scores of the PCL screen, imaged and analysed at EPFL, Processed Plaque-2.0 infection scores of the PCL screen, imaged and analysed at EPFL, respectively<sup><a href="#CR29" class="usa-link" aria-describedby="CR29">29</a></sup>). Both dry-lab pipelines confirmed the high assay quality (Tables <a href="#Tab2" class="usa-link">2</a> and <a href="#Tab3" class="usa-link">3</a>). During hit filtering, PCL compounds that gave significant toxicity in uninfected cells were excluded during hit filtering (Fig. <a href="#Fig5" class="usa-link">5</a>, PCL compounds excluded due to toxicity in uninfected cells<sup><a href="#CR29" class="usa-link" aria-describedby="CR29">29</a></sup>). As summarized in Fig. <a href="#Fig6" class="usa-link">6</a> left panel, both scores are strongly correlated with R<sup>2</sup> between 0.6870 and 0.9870. Both approaches yielded identical top scored compounds (Fig. <a href="#Fig6" class="usa-link">6</a>, right panel), of which Prestw-1764, Nelfinavir mesylate, was the top hit.</p> <figure class="fig xbox font-sm" id="Fig5"><h4 class="obj_head">Fig. 5.</h4> <p class="img-box line-height-none margin-x-neg-2 tablet:margin-x-0 text-center"><img class="graphic" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d5e/7423605/4a09270c25b2/41597_2020_604_Fig5_HTML.jpg" loading="lazy" id="d30e2714" height="149" width="665" alt="Fig. 5"></p> <div class="p text-right font-secondary"><a href="figure/Fig5/" class="usa-link" target="_blank" rel="noopener noreferrer">Open in a new tab</a></div> <figcaption><p>PCL compound toxicity in uninfected cells. Of the 1,278 PCL compounds tested, 126 PCL compounds are found to be toxic, as shown in red, and listed in PCL compounds excluded due to toxicity in uninfected cells<sup><a href="#CR29" class="usa-link" aria-describedby="CR29">29</a></sup>. A549 cells were treated with PCL compounds in duplicates according to the screening wet-lab protocol, however, in absence of HAdV infection for 3.5 days. Doxorubicin hydrochloride (Prestw-438) was used as a positive control for cytotoxicity, at a final concentration of 10 µM, and the corresponding concentration of the drug solvent DMSO was used as a negative control. Cell viability was determined by PrestoBlue assay. PrestoBlue fluorescence intensities of each well were normalized per plate to negative control values at 0 and positive controls at 1. Compounds were considered toxic, when the normalized value for all replicates was higher than the average +3σ (standard deviation, SD) of the DMSO negative control for the corresponding plate. X-axis indicates compounds by their PCL identifier (<em>PCL ID</em>, see PCL compounds tested in the screening procedure<sup><a href="#CR29" class="usa-link" aria-describedby="CR29">29</a></sup>). Normalized average PrestoBlue read-outs are depicted on the y-axis.</p></figcaption></figure><figure class="fig xbox font-sm" id="Fig6"><h4 class="obj_head">Fig. 6.</h4> <p class="img-box line-height-none margin-x-neg-2 tablet:margin-x-0 text-center"><a class="tileshop" target="_blank" href="https://www.ncbi.nlm.nih.gov/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Click%20on%20image%20to%20zoom&p=PMC3&id=7423605_41597_2020_604_Fig6_HTML.jpg"><img class="graphic zoom-in" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d5e/7423605/884068c11b0d/41597_2020_604_Fig6_HTML.jpg" loading="lazy" id="d30e2761" height="1147" width="619" alt="Fig. 6"></a></p> <div class="p text-right font-secondary"><a href="figure/Fig6/" class="usa-link" target="_blank" rel="noopener noreferrer">Open in a new tab</a></div> <figcaption><p>Infection scores from independent dry-lab pipelines. Imaging, image analysis and data processing were performed independently at UZH and EPFL. Infection phenotypes in PCL-treated cells of four biological replicates were averaged and normalized against the DMSO solvent control. Linear regression plots of UZH and EPFL data are shown for (<strong>a</strong>) <em>numberOfNuclei</em>, (<strong>b</strong>) <em>numberOfInfectedNuclei</em>, (<strong>c</strong>) <em>infectionIndex</em>, (<strong>d</strong>) <em>numberOfPlaques</em> and (<strong>e</strong>) <em>totalVirusIntensity</em> of the 1,278 tested PCL compounds from are (green line). Red dots indicate toxicity in the absence of infection. Non-toxic compounds are shown as green dots. R<sup>2</sup> was calculated using GraphPad Prism 8.2.1. Highest scoring compounds are shown on the right, including the <em>PCL_ID</em> of some non-toxic compounds.</p></figcaption></figure></section></section><section id="Sec22"><h2 class="pmc_sec_title">Usage Notes</h2> <p id="Par37">Five parameters were used to score the infection phenotype of each well: the number of nuclei (<em>numberOfNuclei</em>), number of infected nuclei (<em>numberOfInfectedNuclei</em>), the ratio between number of infected and total nuclei (<em>infectionIndex</em>), the number of multi-round infection foci termed plaques (<em>numberOfPlaques</em>) and the extend of viral GFP reporter expression as integrated GFP intensity <em>totalVirusIntensity</em>).</p> <section id="Sec23"><h3 class="pmc_sec_title">Infection scoring using the Plaque2.0 GUI</h3> <p id="Par38">A detailed manual for Plaque2.0 GUI-based infection phenotype scoring is available at <a href="https://plaque2.github.io/" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">plaque2.github.io/</a>. No MATLAB license is necessary.</p> <p id="Par39">The following settings should be used:</p> <p id="Par40"><span class="text-underline">Input/Output</span>:</p> <p id="Par41">Processing folder: Path to folder containing the images (e.g. <em>idr0081/3-Screen/Data_EPFL/Screen/ BSF018292_1A</em>).</p> <p id="Par42"><em>filename pattern</em> Data_UZH:<em>.* (? < wellName > [A-Z][0–9]*)_(? < channelName > w[0–9]*).TIF</em></p> <p id="Par43"><em>filename pattern</em> Data_EPFL:<em>.* (? < wellName > [A-Z] - [0–9] + )[(]fld 1 wv (? < channel > [A-Z]{4}).*.tif</em></p> <p id="Par44"><em>Plate name</em>: Name of the plate to be analysed (e.g. <em>BSF018292_1A</em>)</p> <p id="Par45"><em>Result Output Folder</em>: Path to the results folder in the respective data folder (e.g. <em>idr0081/3-Screen/Data_EPFL/Results</em>).</p> <p id="Par46"><span class="text-underline">Stitch:</span> Stitching of the images is not necessary, since every 384-well is imaged in a single site. Do not activate the tab.</p> <p id="Par47"><span class="text-underline">Mask</span>:</p> <p id="Par48"><em>Custom Mask File</em>: Path to the manually defined mask file (e.g. <em>idr0081/3-Screen/Data_UZH/Parameters</em>). Well masking is optional and was not performed by EPFL.</p> <p id="Par49"><span class="text-underline">Monolayer</span>:</p> <p id="Par50"><em>Channel</em>: Nuclei were imaged in channel 1.</p> <p id="Par51"><span class="text-underline">Plaque</span>:</p> <p id="Par52"><em>Channel</em>: Viral GFP reporter signal was imaged in channel 2.</p></section><section id="Sec24"><h3 class="pmc_sec_title">Infection scoring using the Plaque2.0 batch script</h3> <p id="Par53">How to use the <em>AntiVir_batchprocessing.m</em> for Plaque2.0 batch processing is indicated in the comments of the code.</p></section></section><section id="ack1" class="ack"><h2 class="pmc_sec_title">Acknowledgements</h2> <p>We thank the Greber lab for fruitful discussions and critical assessment of the data. We thank the IDR team for helping to make our work openly accessible. The work was supported by the Swiss National Science Foundation (SNSF) to U.F.G. (Grant numbers 316030_170799/1 and 31003A_179256/1), and the SNSF National Research Program “NCCR Chemical Biology” to G.T. and U.F.G.</p></section><section id="notes1"><h2 class="pmc_sec_title">Author contributions</h2> <p>U.F.G., V.A., A.Y. conceived the screening idea. F.G. designed the experiments, and with U.F.G. coordinated the project. F.K. prepared the PCL-spotted plates. F.G. and R.W. performed the experiments. F.G. and F.K. acquired the data. F.G. and V.A. analysed the imaging data. L.M. and F.G. processed the data. G.T. organized and supervised the screening project at the EPFL-BSF. F.G., F.K. and U.F.G. wrote manuscript, with input from all the co-authors.</p></section><section id="notes2"><h2 class="pmc_sec_title">Code availability</h2> <p><strong>Plaque2.0 batch image analysis for infection scoring</strong>. The MATLAB (version R2016b, The MathWorks, Natick, USA) script <em>AntiVir_batchprocessing.m</em> used by UZH for image analysis is provided for download at IDR, accession number idr0081, under <em>idr0081/3-Screen/Analysis</em>. It is based on the Plaque2.0 software available on GitHub under GPLv3 open source license: <a href="https://github.com/plaque2/matlab" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">https://github.com/plaque2/matlab</a>.</p> <p>To batch analyse the HAdV screening data by Plaque2.0, fork or download the Plaque2.0 AntiVir code from GitHub: <a href="https://github.com/plaque2/matlab/tree/antivir" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">https://github.com/plaque2/matlab/tree/antivir</a>. Place the <em>AntiVir_batchprocessing.m</em> file from <em>idr0081/3-Screen/Analysis</em> into the <em>Plaque2/matlab</em> folder and follow the instructions in <em>AntiVir_batchprocessing.m</em>. A MATLAB license is required.</p> <p><strong>Hit filtering using R</strong>. The R<sup><a href="#CR39" class="usa-link" aria-describedby="CR39">39</a></sup> (version 3.6.1 (2019-07-05)) script <em>AntiVir_hitfiltering.R</em> used by UZH for data processing and hit filtering is provided at IDR accession number idr0081 under <em>idr0081/3-Screen/Analysis</em>.</p></section><section id="FPar1"><h2 class="pmc_sec_title">Competing interests</h2> <p id="Par54">The authors declare no competing interests.</p></section><section id="fn-group1" class="fn-group"><h2 class="pmc_sec_title">Footnotes</h2> <div class="fn-group p font-secondary-light font-sm"><div class="fn p" id="fn1"><p><strong>Publisher’s note</strong> Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.</p></div></div></section><section id="Bib1" class="ref-list"><h2 class="pmc_sec_title">References</h2> <section id="Bib1_sec2"><ul class="ref-list font-sm" style="list-style-type:none"> <li id="CR1"> <span class="label">1.</span><cite>Krilov LR. Adenovirus infections in the immunocompromised host. Pediatr Infect Dis J. 2005;24:555–556. doi: 10.1097/01.inf.0000165306.95815.68.</cite> [<a href="https://doi.org/10.1097/01.inf.0000165306.95815.68" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/15933569/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Pediatr%20Infect%20Dis%20J&title=Adenovirus%20infections%20in%20the%20immunocompromised%20host&author=LR%20Krilov&volume=24&publication_year=2005&pages=555-556&pmid=15933569&doi=10.1097/01.inf.0000165306.95815.68&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR2"> <span class="label">2.</span><cite>Greber UF, Arnberg N, Wadell G, Benkő M, Kremer EJ. Adenoviruses - from pathogens to therapeutics: a report on the 10th International Adenovirus Meeting. Cell Microbiol. 2013;15:16–23. doi: 10.1111/cmi.12031.</cite> [<a href="https://doi.org/10.1111/cmi.12031" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/22985121/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Cell%20Microbiol&title=Adenoviruses%20-%20from%20pathogens%20to%20therapeutics:%20a%20report%20on%20the%2010th%20International%20Adenovirus%20Meeting&author=UF%20Greber&author=N%20Arnberg&author=G%20Wadell&author=M%20Benk%C5%91&author=EJ%20Kremer&volume=15&publication_year=2013&pages=16-23&pmid=22985121&doi=10.1111/cmi.12031&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR3"> <span class="label">3.</span><cite>Tunkel AR, Baron EL, Buch KA, Marty FM, Martinez-Lage M. Case 31-2019: A 45-Year-Old Woman with Headache and Somnolence. N Engl J Med. 2019;381:1459–1470. doi: 10.1056/NEJMcpc1904045.</cite> [<a href="https://doi.org/10.1056/NEJMcpc1904045" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/31597024/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=N%20Engl%20J%20Med&title=Case%2031-2019:%20A%2045-Year-Old%20Woman%20with%20Headache%20and%20Somnolence&author=AR%20Tunkel&author=EL%20Baron&author=KA%20Buch&author=FM%20Marty&author=M%20Martinez-Lage&volume=381&publication_year=2019&pages=1459-1470&pmid=31597024&doi=10.1056/NEJMcpc1904045&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR4"> <span class="label">4.</span><cite>Gray GC, et al. Genotype prevalence and risk factors for severe clinical adenovirus infection, United States 2004-2006. Clin Infect Dis. 2007;45:1120–1131. doi: 10.1086/522188.</cite> [<a href="https://doi.org/10.1086/522188" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC2064001/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/17918073/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Clin%20Infect%20Dis&title=Genotype%20prevalence%20and%20risk%20factors%20for%20severe%20clinical%20adenovirus%20infection,%20United%20States%202004-2006&author=GC%20Gray&volume=45&publication_year=2007&pages=1120-1131&pmid=17918073&doi=10.1086/522188&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR5"> <span class="label">5.</span><cite>Metzgar D, et al. Abrupt emergence of diverse species B adenoviruses at US military recruit training centers. J Infect Dis. 2007;196:1465–1473. doi: 10.1086/522970.</cite> [<a href="https://doi.org/10.1086/522970" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/18008225/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=J%20Infect%20Dis&title=Abrupt%20emergence%20of%20diverse%20species%20B%20adenoviruses%20at%20US%20military%20recruit%20training%20centers&author=D%20Metzgar&volume=196&publication_year=2007&pages=1465-1473&pmid=18008225&doi=10.1086/522970&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR6"> <span class="label">6.</span><cite>Lynch JP, Kajon AE. Adenovirus: epidemiology, global spread of novel serotypes, and advances in treatment and prevention. Semin Respir Crit Care Med. 2016;37:586–602. doi: 10.1055/s-0036-1584923.</cite> [<a href="https://doi.org/10.1055/s-0036-1584923" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC7171713/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/27486739/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Semin%20Respir%20Crit%20Care%20Med&title=Adenovirus:%20epidemiology,%20global%20spread%20of%20novel%20serotypes,%20and%20advances%20in%20treatment%20and%20prevention&author=JP%20Lynch&author=AE%20Kajon&volume=37&publication_year=2016&pages=586-602&pmid=27486739&doi=10.1055/s-0036-1584923&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR7"> <span class="label">7.</span><cite>Haque E, Banik U, Monowar T, Anthony L, Adhikary AK. Worldwide increased prevalence of human adenovirus type 3 (HAdV-3) respiratory infections is well correlated with heterogeneous hypervariable regions (HVRs) of hexon. PLoS ONE. 2018;13:e0194516. doi: 10.1371/journal.pone.0194516.</cite> [<a href="https://doi.org/10.1371/journal.pone.0194516" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC5874027/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/29590206/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=PLoS%20ONE&title=Worldwide%20increased%20prevalence%20of%20human%20adenovirus%20type%203%20(HAdV-3)%20respiratory%20infections%20is%20well%20correlated%20with%20heterogeneous%20hypervariable%20regions%20(HVRs)%20of%20hexon&author=E%20Haque&author=U%20Banik&author=T%20Monowar&author=L%20Anthony&author=AK%20Adhikary&volume=13&publication_year=2018&pages=e0194516&pmid=29590206&doi=10.1371/journal.pone.0194516&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR8"> <span class="label">8.</span><cite>Jiang H, et al. Oncolytic adenovirus research evolution: from cell-cycle checkpoints to immune checkpoints. Curr Opin Virol. 2015;13:33–39. doi: 10.1016/j.coviro.2015.03.009.</cite> [<a href="https://doi.org/10.1016/j.coviro.2015.03.009" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC4550502/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/25863716/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Curr%20Opin%20Virol&title=Oncolytic%20adenovirus%20research%20evolution:%20from%20cell-cycle%20checkpoints%20to%20immune%20checkpoints&author=H%20Jiang&volume=13&publication_year=2015&pages=33-39&pmid=25863716&doi=10.1016/j.coviro.2015.03.009&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR9"> <span class="label">9.</span><cite>Lawler SE, Speranza M-C, Cho C-F, Chiocca EA. Oncolytic viruses in cancer treatment: A review. JAMA Oncology. 2017;3:841–849. doi: 10.1001/jamaoncol.2016.2064.</cite> [<a href="https://doi.org/10.1001/jamaoncol.2016.2064" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/27441411/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=JAMA%20Oncology&title=Oncolytic%20viruses%20in%20cancer%20treatment:%20A%20review&author=SE%20Lawler&author=M-C%20Speranza&author=C-F%20Cho&author=EA%20Chiocca&volume=3&publication_year=2017&pages=841-849&pmid=27441411&doi=10.1001/jamaoncol.2016.2064&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR10"> <span class="label">10.</span><cite>Ginn SL, Amaya AK, Alexander IE, Edelstein M, Abedi MR. Gene therapy clinical trials worldwide to 2017: An update. J Gene Med. 2018;20:e3015. doi: 10.1002/jgm.3015.</cite> [<a href="https://doi.org/10.1002/jgm.3015" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/29575374/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=J%20Gene%20Med&title=Gene%20therapy%20clinical%20trials%20worldwide%20to%202017:%20An%20update&author=SL%20Ginn&author=AK%20Amaya&author=IE%20Alexander&author=M%20Edelstein&author=MR%20Abedi&volume=20&publication_year=2018&pages=e3015&pmid=29575374&doi=10.1002/jgm.3015&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR11"> <span class="label">11.</span><cite>Mennechet FJD, et al. A review of 65 years of human adenovirus seroprevalence. Expert Rev Vaccines. 2019;18:597–613. doi: 10.1080/14760584.2019.1588113.</cite> [<a href="https://doi.org/10.1080/14760584.2019.1588113" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/31132024/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Expert%20Rev%20Vaccines&title=A%20review%20of%2065%20years%20of%20human%20adenovirus%20seroprevalence&author=FJD%20Mennechet&volume=18&publication_year=2019&pages=597-613&pmid=31132024&doi=10.1080/14760584.2019.1588113&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR12"> <span class="label">12.</span><cite>Lion, T. Adenovirus persistence, reactivation, and clinical management. <em>FEBS Lett</em>, 10.1002/1873-3468.13576 (2019).</cite> [<a href="https://doi.org/10.1002/1873-3468.13576" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/31411731/" class="usa-link">PubMed</a>]</li> <li id="CR13"> <span class="label">13.</span><cite>Ismail AM, et al. Genomic foundations of evolution and ocular pathogenesis in human adenovirus species D. FEBS Lett. 2019;593:3583–3608. doi: 10.1002/1873-3468.13693.</cite> [<a href="https://doi.org/10.1002/1873-3468.13693" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC7185199/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/31769017/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=FEBS%20Lett&title=Genomic%20foundations%20of%20evolution%20and%20ocular%20pathogenesis%20in%20human%20adenovirus%20species%20D&author=AM%20Ismail&volume=593&publication_year=2019&pages=3583-3608&pmid=31769017&doi=10.1002/1873-3468.13693&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR14"> <span class="label">14.</span><cite>Harrach, B., Tarján, Z. L. & Benkő, M. Adenoviruses across the animal kingdom: a walk in the zoo. <em>FEBS Lett</em>, 10.1002/1873-3468.13687 (2019).</cite> [<a href="https://doi.org/10.1002/1873-3468.13687" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/31747467/" class="usa-link">PubMed</a>]</li> <li id="CR15"> <span class="label">15.</span><cite>Reddy VS, Natchiar SK, Stewart PL, Nemerow GR. Crystal structure of human adenovirus at 3.5 A resolution. Science. 2010;329:1071–1075. doi: 10.1126/science.1187292.</cite> [<a href="https://doi.org/10.1126/science.1187292" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC2929978/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/20798318/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Science&title=Crystal%20structure%20of%20human%20adenovirus%20at%203.5%20A%20resolution&author=VS%20Reddy&author=SK%20Natchiar&author=PL%20Stewart&author=GR%20Nemerow&volume=329&publication_year=2010&pages=1071-1075&pmid=20798318&doi=10.1126/science.1187292&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR16"> <span class="label">16.</span><cite>Benevento M, et al. Adenovirus composition, proteolysis, and disassembly studied by in-depth qualitative and quantitative proteomics. J Biol Chem. 2014;289:11421–11430. doi: 10.1074/jbc.M113.537498.</cite> [<a href="https://doi.org/10.1074/jbc.M113.537498" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC4036278/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/24591515/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=J%20Biol%20Chem&title=Adenovirus%20composition,%20proteolysis,%20and%20disassembly%20studied%20by%20in-depth%20qualitative%20and%20quantitative%20proteomics&author=M%20Benevento&volume=289&publication_year=2014&pages=11421-11430&pmid=24591515&doi=10.1074/jbc.M113.537498&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR17"> <span class="label">17.</span><cite>Greber UF, Flatt JW. Adenovirus entry: from infection to immunity. Annual review of virology. 2019;6:177–197. doi: 10.1146/annurev-virology-092818-015550.</cite> [<a href="https://doi.org/10.1146/annurev-virology-092818-015550" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/31283442/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Annual%20review%20of%20virology&title=Adenovirus%20entry:%20from%20infection%20to%20immunity&author=UF%20Greber&author=JW%20Flatt&volume=6&publication_year=2019&pages=177-197&pmid=31283442&doi=10.1146/annurev-virology-092818-015550&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR18"> <span class="label">18.</span><cite>Bauer M, et al. The e3 ubiquitin ligase mind bomb 1 controls adenovirus genome release at the nuclear pore complex. Cell Rep. 2019;29:3785–3795.e8. doi: 10.1016/j.celrep.2019.11.064.</cite> [<a href="https://doi.org/10.1016/j.celrep.2019.11.064" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/31851912/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Cell%20Rep&title=The%20e3%20ubiquitin%20ligase%20mind%20bomb%201%20controls%20adenovirus%20genome%20release%20at%20the%20nuclear%20pore%20complex&author=M%20Bauer&volume=29&publication_year=2019&pages=3785-3795.e8&pmid=31851912&doi=10.1016/j.celrep.2019.11.064&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR19"> <span class="label">19.</span><cite>Greber UF. Virus and host mechanics support membrane penetration and cell entry. J Virol. 2016;90:3802–3805. doi: 10.1128/JVI.02568-15.</cite> [<a href="https://doi.org/10.1128/JVI.02568-15" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC4810568/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/26842477/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=J%20Virol&title=Virus%20and%20host%20mechanics%20support%20membrane%20penetration%20and%20cell%20entry&author=UF%20Greber&volume=90&publication_year=2016&pages=3802-3805&pmid=26842477&doi=10.1128/JVI.02568-15&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR20"> <span class="label">20.</span><cite>Wang I-H, Burckhardt CJ, Yakimovich A, Morf MK, Greber UF. The nuclear export factor CRM1 controls juxta-nuclear microtubule-dependent virus transport. J Cell Sci. 2017;130:2185–2195. doi: 10.1242/jcs.203794.</cite> [<a href="https://doi.org/10.1242/jcs.203794" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/28515232/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=J%20Cell%20Sci&title=The%20nuclear%20export%20factor%20CRM1%20controls%20juxta-nuclear%20microtubule-dependent%20virus%20transport&author=I-H%20Wang&author=CJ%20Burckhardt&author=A%20Yakimovich&author=MK%20Morf&author=UF%20Greber&volume=130&publication_year=2017&pages=2185-2195&pmid=28515232&doi=10.1242/jcs.203794&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR21"> <span class="label">21.</span><cite>Prasad V, et al. The UPR sensor IRE1α and the adenovirus E3-19K glycoprotein sustain persistent and lytic infections. Nat Commun. 2020;11:1997. doi: 10.1038/s41467-020-15844-2.</cite> [<a href="https://doi.org/10.1038/s41467-020-15844-2" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC7181865/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/32332742/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Nat%20Commun&title=The%20UPR%20sensor%20IRE1%CE%B1%20and%20the%20adenovirus%20E3-19K%20glycoprotein%20sustain%20persistent%20and%20lytic%20infections&author=V%20Prasad&volume=11&publication_year=2020&pages=1997&pmid=32332742&doi=10.1038/s41467-020-15844-2&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR22"> <span class="label">22.</span><cite>King, C. R., Zhang, A., Tessier, T. M., Gameiro, S. F. & Mymryk, J. S. Hacking the cell: network intrusion and exploitation by adenovirus E1A. <em>MBio</em><strong>9</strong>, (2018).</cite> [<a href="https://doi.org/10.1128/mBio.00390-18" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC5930299/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/29717008/" class="usa-link">PubMed</a>]</li> <li id="CR23"> <span class="label">23.</span><cite>Zheng Y, Stamminger T, Hearing P. E2f/rb family proteins mediate interferon induced repression of adenovirus immediate early transcription to promote persistent viral infection. PLoS Pathog. 2016;12:e1005415. doi: 10.1371/journal.ppat.1005415.</cite> [<a href="https://doi.org/10.1371/journal.ppat.1005415" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC4726734/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/26809031/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=PLoS%20Pathog&title=E2f/rb%20family%20proteins%20mediate%20interferon%20induced%20repression%20of%20adenovirus%20immediate%20early%20transcription%20to%20promote%20persistent%20viral%20infection&author=Y%20Zheng&author=T%20Stamminger&author=P%20Hearing&volume=12&publication_year=2016&pages=e1005415&pmid=26809031&doi=10.1371/journal.ppat.1005415&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR24"> <span class="label">24.</span><cite>Yakimovich A, et al. Cell-free transmission of human adenovirus by passive mass transfer in cell culture simulated in a computer model. J Virol. 2012;86:10123–10137. doi: 10.1128/JVI.01102-12.</cite> [<a href="https://doi.org/10.1128/JVI.01102-12" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC3446567/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/22787215/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=J%20Virol&title=Cell-free%20transmission%20of%20human%20adenovirus%20by%20passive%20mass%20transfer%20in%20cell%20culture%20simulated%20in%20a%20computer%20model&author=A%20Yakimovich&volume=86&publication_year=2012&pages=10123-10137&pmid=22787215&doi=10.1128/JVI.01102-12&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR25"> <span class="label">25.</span><cite>Tollefson AE, et al. The adenovirus death protein (E3-11.6K) is required at very late stages of infection for efficient cell lysis and release of adenovirus from infected cells. J Virol. 1996;70:2296–2306. doi: 10.1128/JVI.70.4.2296-2306.1996.</cite> [<a href="https://doi.org/10.1128/JVI.70.4.2296-2306.1996" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC190071/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/8642656/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=J%20Virol&title=The%20adenovirus%20death%20protein%20(E3-11.6K)%20is%20required%20at%20very%20late%20stages%20of%20infection%20for%20efficient%20cell%20lysis%20and%20release%20of%20adenovirus%20from%20infected%20cells&author=AE%20Tollefson&volume=70&publication_year=1996&pages=2296-2306&pmid=8642656&doi=10.1128/JVI.70.4.2296-2306.1996&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR26"> <span class="label">26.</span><cite>Doronin K, et al. Overexpression of the ADP (E3-11.6K) protein increases cell lysis and spread of adenovirus. Virology. 2003;305:378–387. doi: 10.1006/viro.2002.1772.</cite> [<a href="https://doi.org/10.1006/viro.2002.1772" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/12573583/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Virology&title=Overexpression%20of%20the%20ADP%20(E3-11.6K)%20protein%20increases%20cell%20lysis%20and%20spread%20of%20adenovirus&author=K%20Doronin&volume=305&publication_year=2003&pages=378-387&pmid=12573583&doi=10.1006/viro.2002.1772&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR27"> <span class="label">27.</span><cite>Lenaerts L, Naesens L. Antiviral therapy for adenovirus infections. Antiviral Res. 2006;71:172–180. doi: 10.1016/j.antiviral.2006.04.007.</cite> [<a href="https://doi.org/10.1016/j.antiviral.2006.04.007" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/16698093/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Antiviral%20Res&title=Antiviral%20therapy%20for%20adenovirus%20infections&author=L%20Lenaerts&author=L%20Naesens&volume=71&publication_year=2006&pages=172-180&pmid=16698093&doi=10.1016/j.antiviral.2006.04.007&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR28"> <span class="label">28.</span><cite>Wold WSM, Tollefson AE, Ying B, Spencer JF, Toth K. Drug development against human adenoviruses and its advancement by Syrian hamster models. FEMS Microbiol Rev. 2019;43:380–388. doi: 10.1093/femsre/fuz008.</cite> [<a href="https://doi.org/10.1093/femsre/fuz008" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/30916746/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=FEMS%20Microbiol%20Rev&title=Drug%20development%20against%20human%20adenoviruses%20and%20its%20advancement%20by%20Syrian%20hamster%20models&author=WSM%20Wold&author=AE%20Tollefson&author=B%20Ying&author=JF%20Spencer&author=K%20Toth&volume=43&publication_year=2019&pages=380-388&pmid=30916746&doi=10.1093/femsre/fuz008&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR29"> <span class="label">29.</span><cite>Georgi F, 2020. High-content image-based drug screen identifies a clinical compound against cell transmission of adenovirus. figshare. </cite> [<a href="https://doi.org/10.6084/m9.figshare.c.5052965" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC7423605/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/32788590/" class="usa-link">PubMed</a>]</li> <li id="CR30"> <span class="label">30.</span><cite>Chauvin C, et al. High-Throughput Drug Screening Identifies Pazopanib and Clofilium Tosylate as Promising Treatments for Malignant Rhabdoid Tumors. Cell Rep. 2017;21:1737–1745. doi: 10.1016/j.celrep.2017.10.076.</cite> [<a href="https://doi.org/10.1016/j.celrep.2017.10.076" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/29141209/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Cell%20Rep&title=High-Throughput%20Drug%20Screening%20Identifies%20Pazopanib%20and%20Clofilium%20Tosylate%20as%20Promising%20Treatments%20for%20Malignant%20Rhabdoid%20Tumors&author=C%20Chauvin&volume=21&publication_year=2017&pages=1737-1745&pmid=29141209&doi=10.1016/j.celrep.2017.10.076&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR31"> <span class="label">31.</span><cite>Wall, G. <em>et al</em>. Screening a Repurposing Library for Inhibitors of Multidrug-Resistant Candida auris Identifies Ebselen as a Repositionable Candidate for Antifungal Drug Development. <em>Antimicrob Agents Chemother</em><strong>62</strong> (2018).</cite> [<a href="https://doi.org/10.1128/AAC.01084-18" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC6153848/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/30104269/" class="usa-link">PubMed</a>]</li> <li id="CR32"> <span class="label">32.</span><cite>Yakimovich A, et al. Plaque2.0-A High-Throughput Analysis Framework to Score Virus-Cell Transmission and Clonal Cell Expansion. PLoS ONE. 2015;10:e0138760. doi: 10.1371/journal.pone.0138760.</cite> [<a href="https://doi.org/10.1371/journal.pone.0138760" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC4587671/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/26413745/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=PLoS%20ONE&title=Plaque2.0-A%20High-Throughput%20Analysis%20Framework%20to%20Score%20Virus-Cell%20Transmission%20and%20Clonal%20Cell%20Expansion&author=A%20Yakimovich&volume=10&publication_year=2015&pages=e0138760&pmid=26413745&doi=10.1371/journal.pone.0138760&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR33"> <span class="label">33.</span><cite>Georgi F, 2020. High-content image-based drug screen identifies a clinical compound against cell transmission of adenovirus. University of Dundee. </cite> [<a href="https://doi.org/10.17867/10000136" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC7423605/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/32788590/" class="usa-link">PubMed</a>]</li> <li id="CR34"> <span class="label">34.</span><cite>Georgi, F. <em>et al</em>. The FDA-approved drug Nelfinavir inhibits lytic cell-free, but not cell-associated non-lytic transmission of human adenovirus. <em>Antimicrob Agents Chemother</em>, 10.1128/AAC.01002-20 (2020).</cite> [<a href="https://doi.org/10.1128/AAC.01002-20" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC7449217/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/32601166/" class="usa-link">PubMed</a>]</li> <li id="CR35"> <span class="label">35.</span><cite>Georgi, F. <em>et al</em>. Mutant Human adenovirus 2 isolate HAdV-C2-dE3B-CMV-GFP, complete sequence. <em>GenBank</em><a href="https://identifiers.org/ncbi/insdc:MT277585.1" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">https://identifiers.org/ncbi/insdc:MT277585.1</a> (2020).</cite> </li> <li id="CR36"> <span class="label">36.</span><cite>Greber UF, Willetts M, Webster P, Helenius A. Stepwise dismantling of adenovirus 2 during entry into cells. Cell. 1993;75:477–486. doi: 10.1016/0092-8674(93)90382-Z.</cite> [<a href="https://doi.org/10.1016/0092-8674(93)90382-Z" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/8221887/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=Cell&title=Stepwise%20dismantling%20of%20adenovirus%202%20during%20entry%20into%20cells&author=UF%20Greber&author=M%20Willetts&author=P%20Webster&author=A%20Helenius&volume=75&publication_year=1993&pages=477-486&pmid=8221887&doi=10.1016/0092-8674(93)90382-Z&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR37"> <span class="label">37.</span><cite>Boncler M, Różalski M, Krajewska U, Podsędek A, Watala C. Comparison of PrestoBlue and MTT assays of cellular viability in the assessment of anti-proliferative effects of plant extracts on human endothelial cells. J Pharmacol Toxicol Methods. 2014;69:9–16. doi: 10.1016/j.vascn.2013.09.003.</cite> [<a href="https://doi.org/10.1016/j.vascn.2013.09.003" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/24103906/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=J%20Pharmacol%20Toxicol%20Methods&title=Comparison%20of%20PrestoBlue%20and%20MTT%20assays%20of%20cellular%20viability%20in%20the%20assessment%20of%20anti-proliferative%20effects%20of%20plant%20extracts%20on%20human%20endothelial%20cells&author=M%20Boncler&author=M%20R%C3%B3%C5%BCalski&author=U%20Krajewska&author=A%20Pods%C4%99dek&author=C%20Watala&volume=69&publication_year=2014&pages=9-16&pmid=24103906&doi=10.1016/j.vascn.2013.09.003&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR38"> <span class="label">38.</span><cite>Xu M, McCanna DJ, Sivak JG. Use of the viability reagent PrestoBlue in comparison with alamarBlue and MTT to assess the viability of human corneal epithelial cells. J Pharmacol Toxicol Methods. 2015;71:1–7. doi: 10.1016/j.vascn.2014.11.003.</cite> [<a href="https://doi.org/10.1016/j.vascn.2014.11.003" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/25464019/" class="usa-link">PubMed</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=J%20Pharmacol%20Toxicol%20Methods&title=Use%20of%20the%20viability%20reagent%20PrestoBlue%20in%20comparison%20with%20alamarBlue%20and%20MTT%20to%20assess%20the%20viability%20of%20human%20corneal%20epithelial%20cells&author=M%20Xu&author=DJ%20McCanna&author=JG%20Sivak&volume=71&publication_year=2015&pages=1-7&pmid=25464019&doi=10.1016/j.vascn.2014.11.003&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> <li id="CR39"> <span class="label">39.</span><cite>R Core Team. <em>R: A Language and Environment for Statistical Computing</em>. (R Foundation for Statistical Computing, 2018).</cite> </li> <li id="CR40"> <span class="label">40.</span><cite>Berthold MR, et al. KNIME - the Konstanz information miner. ACM SIGKDD Explorations Newsletter. 2009;11:26. doi: 10.1145/1656274.1656280.</cite> [<a href="https://doi.org/10.1145/1656274.1656280" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="https://scholar.google.com/scholar_lookup?journal=ACM%20SIGKDD%20Explorations%20Newsletter&title=KNIME%20-%20the%20Konstanz%20information%20miner&author=MR%20Berthold&volume=11&publication_year=2009&pages=26&doi=10.1145/1656274.1656280&" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">Google Scholar</a>]</li> </ul></section></section><section id="_ad93_" lang="en" class="associated-data"><h2 class="pmc_sec_title">Associated Data</h2> <p class="font-secondary"><em>This section collects any data citations, data availability statements, or supplementary materials included in this article.</em></p> <section id="_adc93_" lang="en" class="data-citations"><h3 class="pmc_sec_title">Data Citations</h3> <ol class="ref-list font-sm"> <li id="_dbx_ref_element-citation26"> <cite>Georgi F, 2020. High-content image-based drug screen identifies a clinical compound against cell transmission of adenovirus. figshare. </cite> [<a href="https://doi.org/10.6084/m9.figshare.c.5052965" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC7423605/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/32788590/" class="usa-link">PubMed</a>]</li> <li id="_dbx_ref_element-citation29"> <cite>Georgi F, 2020. High-content image-based drug screen identifies a clinical compound against cell transmission of adenovirus. University of Dundee. </cite> [<a href="https://doi.org/10.17867/10000136" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">DOI</a>] [<a href="/articles/PMC7423605/" class="usa-link">PMC free article</a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/32788590/" class="usa-link">PubMed</a>]</li> </ol></section><section id="_adda93_" lang="en" class="data-availability-statement"><h3 class="pmc_sec_title">Data Availability Statement</h3> <p><strong>Plaque2.0 batch image analysis for infection scoring</strong>. The MATLAB (version R2016b, The MathWorks, Natick, USA) script <em>AntiVir_batchprocessing.m</em> used by UZH for image analysis is provided for download at IDR, accession number idr0081, under <em>idr0081/3-Screen/Analysis</em>. It is based on the Plaque2.0 software available on GitHub under GPLv3 open source license: <a href="https://github.com/plaque2/matlab" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">https://github.com/plaque2/matlab</a>.</p> <p>To batch analyse the HAdV screening data by Plaque2.0, fork or download the Plaque2.0 AntiVir code from GitHub: <a href="https://github.com/plaque2/matlab/tree/antivir" class="usa-link usa-link--external" data-ga-action="click_feat_suppl" target="_blank" rel="noopener noreferrer">https://github.com/plaque2/matlab/tree/antivir</a>. Place the <em>AntiVir_batchprocessing.m</em> file from <em>idr0081/3-Screen/Analysis</em> into the <em>Plaque2/matlab</em> folder and follow the instructions in <em>AntiVir_batchprocessing.m</em>. A MATLAB license is required.</p> <p><strong>Hit filtering using R</strong>. The R<sup><a href="#CR39" class="usa-link" aria-describedby="CR39">39</a></sup> (version 3.6.1 (2019-07-05)) script <em>AntiVir_hitfiltering.R</em> used by UZH for data processing and hit filtering is provided at IDR accession number idr0081 under <em>idr0081/3-Screen/Analysis</em>.</p></section></section></section><footer class="p courtesy-note font-secondary font-sm text-center"><hr class="headless"> <p>Articles from Scientific Data are provided here courtesy of <strong>Nature Publishing Group</strong></p></footer></section></article> </main> </div> </div> </div>  <div class="pmc-sidenav desktop:grid-col-4 display-flex"> <section class="pmc-sidenav__container" aria-label="Article resources and navigation"> <button type="button" class="usa-button pmc-sidenav__container__close usa-button--unstyled"> <img src="/static/img/usa-icons/close.svg" role="img" alt="Close" /> </button> <div class="display-none desktop:display-block"> <section class="margin-top-4 desktop:margin-top-0"> <h2 class="margin-top-0">ACTIONS</h2> <ul class="usa-list usa-list--unstyled usa-list--actions"> <li> <a href="https://doi.org/10.1038/s41597-020-00604-0" class="usa-button usa-button--outline width-24 font-xs usa-link--external padding-left-0 padding-right-0" target="_blank" rel="noreferrer noopener" data-ga-category="actions" data-ga-action="click" data-ga-label="publisher_link_desktop" > <span class="height-3 display-inline-flex flex-align-center">View on publisher site</span> </a> </li> <li> <a href="pdf/41597_2020_Article_604.pdf" class="usa-button usa-button--outline width-24 display-inline-flex flex-align-center flex-justify-start padding-left-1" data-ga-category="actions" data-ga-action="click" data-ga-label="pdf_download_desktop" > <svg class="usa-icon width-3 height-3" aria-hidden="true" focusable="false" role="img" hidden> <use xlink:href="/static/img/sprite.svg#file_download"></use> </svg> <span class="display-inline-flex flex-justify-center flex-1">PDF (2.8 MB)</span> </a> </li> <li> <button role="button" class="usa-button width-24 citation-dialog-trigger display-inline-flex flex-align-center flex-justify-start padding-left-1" aria-label="Open dialog with citation text in different styles" data-ga-category="actions" data-ga-action="open" data-ga-label="cite_desktop" data-all-citations-url="/resources/citations/7423605/" data-citation-style="nlm" data-download-format-link="/resources/citations/7423605/export/" > <svg class="usa-icon width-3 height-3" aria-hidden="true" focusable="false" role="img" hidden> <use xlink:href="/static/img/sprite.svg#format_quote"></use> </svg> <span class="display-inline-flex flex-justify-center flex-1 button-label">Cite</span> </button> </li> <li> <button class="usa-button width-24 collections-dialog-trigger collections-button display-inline-flex flex-align-center flex-justify-start padding-left-1 collections-button-empty" aria-label="Save article in MyNCBI collections." data-ga-category="actions" data-ga-action="click" data-ga-label="collections_button_desktop" data-collections-open-dialog-enabled="false" data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpmc.ncbi.nlm.nih.gov%2Farticles%2FPMC7423605%2F%23open-collections-dialog" data-in-collections="false"> <svg class="usa-icon width-3 height-3 usa-icon--bookmark-full" aria-hidden="true" focusable="false" role="img" hidden> <use xlink:href="/static/img/action-bookmark-full.svg#icon"></use> </svg> <svg class="usa-icon width-3 height-3 usa-icon--bookmark-empty" aria-hidden="true" focusable="false" role="img" hidden> <use xlink:href="/static/img/action-bookmark-empty.svg#icon"></use> </svg> <span class="display-inline-flex flex-justify-center flex-1">Collections</span> </button> </li> <li class="pmc-permalink"> <button type="button" class="usa-button usa-button--outline width-24 display-inline-flex flex-align-center flex-justify padding-left-1 shadow-none" aria-label="Show article permalink" aria-expanded="false" aria-haspopup="true" data-ga-category="actions" data-ga-action="open" data-ga-label="permalink_desktop" > <svg class="usa-icon width-3 height-3" aria-hidden="true" focusable="false" role="img" hidden> <use xlink:href="/static/img/sprite.svg#share"></use> </svg> <span class="display-inline-flex flex-justify-center flex-1 button-label">Permalink</span> </button> <div class="pmc-permalink__dropdown" hidden> <div class="pmc-permalink__dropdown__container"> <h2 class="usa-modal__heading margin-top-0 margin-bottom-2">PERMALINK</h2> <div class="pmc-permalink__dropdown__content"> <input type="text" class="usa-input" value="https://pmc.ncbi.nlm.nih.gov/articles/PMC7423605/" aria-label="Article permalink"> <button class="usa-button display-inline-flex pmc-permalink__dropdown__copy__btn margin-right-0" title="Copy article permalink" data-ga-category="save_share" data-ga-action="link" data-ga-label="copy_link"> <svg class="usa-icon" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#content_copy"></use> </svg> <span class="margin-left-1">Copy</span> </button> </div> </div> </div> </li> </ul> </section> </div> <section class="pmc-resources margin-top-6 desktop:margin-top-4" data-page-path="/articles/PMC7423605/"> <h2 class="margin-top-0">RESOURCES</h2> <div class="usa-accordion usa-accordion--multiselectable" data-allow-multiple> <h3 class="usa-accordion__heading"> <button type="button" class="usa-accordion__button" aria-expanded="false" aria-controls="resources-similar-articles" data-ga-category="resources_accordion" data-ga-action="open_similar_articles" data-ga-label="/articles/PMC7423605/" data-action-open="open_similar_articles" data-action-close="close_similar_articles" > Similar articles </button> </h3> <div id="resources-similar-articles" class="usa-accordion__content usa-prose" data-source-url="/resources/similar-article-links/32788590/" > </div> <h3 class="usa-accordion__heading"> <button type="button" class="usa-accordion__button" aria-expanded="false" aria-controls="resources-cited-by-other-articles" data-ga-category="resources_accordion" data-ga-action="open_cited_by" data-ga-label="/articles/PMC7423605/" data-action-open="open_cited_by" data-action-close="close_cited_by" > Cited by other articles </button> </h3> <div id="resources-cited-by-other-articles" class="usa-accordion__content usa-prose" data-source-url="/resources/cited-by-links/32788590/" > </div> <h3 class="usa-accordion__heading"> <button type="button" class="usa-accordion__button" aria-expanded="false" aria-controls="resources-links-to-ncbi-databases" data-ga-category="resources_accordion" data-ga-action="open_NCBI_links" data-ga-label="/articles/PMC7423605/" data-action-open="open_NCBI_links" data-action-close="close_NCBI_link" > Links to NCBI Databases </button> </h3> <div id="resources-links-to-ncbi-databases" class="usa-accordion__content usa-prose" data-source-url="/resources/db-links/7423605/" > </div> </div> </section> <section class="usa-in-page-nav usa-in-page-nav--wide margin-top-6 desktop:margin-top-4" data-title-text="On this page" data-title-heading-level="h2" data-scroll-offset="0" data-root-margin="-10% 0px -80% 0px" data-main-content-selector="main" data-threshold="1" hidden ></section> </section> </div> <div class="overlay" role="dialog" aria-label="Citation Dialog" hidden> <div class="dialog citation-dialog" aria-hidden="true"> <div class="display-inline-flex flex-align-center flex-justify width-full margin-bottom-2"> <h2 class="usa-modal__heading margin-0">Cite</h2> <button type="button" class="usa-button usa-button--unstyled close-overlay text-black width-auto" tabindex="1"> <svg class="usa-icon width-3 height-3" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#close"></use> </svg> </button> </div> <div class="citation-text-block"> <div class="citation-text margin-bottom-2"></div> <ul class="usa-list usa-list--unstyled display-inline-flex flex-justify width-full flex-align-center"> <li> <button class="usa-button usa-button--unstyled text-no-underline display-flex flex-align-center copy-button dialog-focus" data-ga-category="save_share" data-ga-action="cite" data-ga-label="copy" tabindex="2"> <svg class="usa-icon width-3 height-3" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#content_copy"></use> </svg> <span>Copy</span> </button> </li> <li> <a href="#" role="button" class="usa-button usa-button--unstyled text-no-underline display-flex flex-align-center export-button" data-ga-category="save_share" data-ga-action="cite" data-ga-label="download" title="Download a file for external citation management software" tabindex="3"> <svg class="usa-icon width-3 height-3" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#file_download"></use> </svg> <span class="display-none mobile-lg:display-inline">Download .nbib</span> <span class="display-inline mobile-lg:display-none">.nbib</span> </a> </li> <li> <div class="display-inline-flex flex-align-center"> <label class="usa-label margin-top-0">Format:</label> <select aria-label="Format" class="usa-select citation-style-selector padding-1 margin-top-0 border-0 padding-right-4" tabindex="4" > <option data-style-url-name="ama" value="AMA" > AMA </option> <option data-style-url-name="apa" value="APA" > APA </option> <option data-style-url-name="mla" value="MLA" > MLA </option> <option data-style-url-name="nlm" value="NLM" selected="selected"> NLM </option> </select> </div> </li> </ul> </div> </div> </div> <div class="overlay" role="dialog" hidden> <div id="collections-action-dialog" class="dialog collections-dialog" aria-hidden="true"> <div class="display-inline-flex flex-align-center flex-justify width-full margin-bottom-2"> <h2 class="usa-modal__heading margin-0">Add to Collections</h2> </div> <div class="collections-action-panel action-panel"> <form id="collections-action-dialog-form" class="usa-form maxw-full collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors" data-existing-collections-url="/list-existing-collections/" data-add-to-existing-collection-url="/add-to-existing-collection/" data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/" data-myncbi-max-collection-name-length="100" data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"> <input type="hidden" name="csrfmiddlewaretoken" value="QFS0OauDq4EFLHbV4XpPLOu2SvvWllmbylQbvFKb5zr2PphCTENBTczlhNoL68Kc"> <fieldset class="usa-fieldset margin-bottom-2"> <div class="usa-radio"> <input type="radio" id="collections-action-dialog-new" class="usa-radio__input usa-radio__input--tile collections-new margin-top-0" name="collections" value="new" data-ga-category="collections_button" data-ga-action="click" data-ga-label="collections_radio_new" /> <label class="usa-radio__label" for="collections-action-dialog-new">Create a new collection</label> </div> <div class="usa-radio"> <input type="radio" id="collections-action-dialog-existing" class="usa-radio__input usa-radio__input--tile collections-existing" name="collections" value="existing" checked="true" data-ga-category="collections_button" data-ga-action="click" data-ga-label="collections_radio_existing" /> <label class="usa-radio__label" for="collections-action-dialog-existing">Add to an existing collection</label> </div> </fieldset> <fieldset class="usa-fieldset margin-bottom-2"> <div class="action-panel-control-wrap new-collections-controls"> <label for="collections-action-dialog-add-to-new" class="usa-label margin-top-0"> Name your collection <abbr title="required" class="usa-hint usa-hint--required text-no-underline">*</abbr> </label> <input type="text" name="add-to-new-collection" id="collections-action-dialog-add-to-new" class="usa-input collections-action-add-to-new" pattern="[^"&=<>/]*" title="The following characters are not allowed in the Name field: "&=<>/" maxlength="" data-ga-category="collections_button" data-ga-action="create_collection" data-ga-label="non_favorties_collection" required /> </div> <div class="action-panel-control-wrap existing-collections-controls"> <label for="collections-action-dialog-add-to-existing" class="usa-label margin-top-0"> Choose a collection </label> <select id="collections-action-dialog-add-to-existing" class="usa-select collections-action-add-to-existing" data-ga-category="collections_button" data-ga-action="select_collection" data-ga-label="($('.collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'"> </select> <div class="collections-retry-load-on-error usa-input-error-message selection-validation-message"> Unable to load your collection due to an error<br> <a href="#">Please try again</a> </div> </div> </fieldset> <div class="display-inline-flex"> <button class="usa-button margin-top-0 action-panel-submit" type="submit" data-loading-label="Adding..." data-pinger-ignore data-ga-category="collections_button" data-ga-action="click" data-ga-label="add"> Add </button> <button class="usa-button usa-button--outline margin-top-0 action-panel-cancel" aria-label="Close 'Add to Collections' panel" ref="linksrc=close_collections_panel" data-ga-category="collections_button" data-ga-action="click" data-ga-label="cancel"> Cancel </button> </div> </form> </div> </div> </div> </div> </div> </div> <footer class="ncbi-footer ncbi-dark-background " > <div class="ncbi-footer__icon-section"> <div class="ncbi-footer__social-header"> Follow NCBI </div> <div class="grid-container ncbi-footer__ncbi-social-icons-container"> <a href="https://twitter.com/ncbi" class="ncbi-footer__social-icon ncbi-footer__social-icon--gray" target="_blank" rel="noreferrer noopener"> <svg width="40" height="40" viewBox="0 0 40 40" fill="none" xmlns="http://www.w3.org/2000/svg" focusable="false" aria-hidden="true"> <path d="m6.067 8 10.81 13.9L6 33.2h4.2l8.4-9.1 7.068 9.1H34L22.8 18.5 31.9 8h-3.5l-7.7 8.4L14.401 8H6.067Zm3.6 1.734h3.266l16.8 21.732H26.57L9.668 9.734Z"> </path> </svg> <span class="usa-sr-only">NCBI on X (formerly known as Twitter)</span> </a> <a href="https://www.facebook.com/ncbi.nlm" class="ncbi-footer__social-icon ncbi-footer__social-icon--gray" target="_blank" rel="noreferrer noopener"> <svg width="16" height="29" focusable="false" aria-hidden="true" viewBox="0 0 16 29" fill="none" xmlns="http://www.w3.org/2000/svg"> <path d="M3.8809 21.4002C3.8809 19.0932 3.8809 16.7876 3.8809 14.478C3.8809 14.2117 3.80103 14.1452 3.54278 14.1492C2.53372 14.1638 1.52334 14.1492 0.514288 14.1598C0.302626 14.1598 0.248047 14.0972 0.248047 13.8936C0.256034 12.4585 0.256034 11.0239 0.248047 9.58978C0.248047 9.37013 0.302626 9.30224 0.528931 9.3049C1.53798 9.31688 2.54837 9.3049 3.55742 9.31555C3.80103 9.31555 3.8809 9.26097 3.87957 9.00272C3.87158 8.00565 3.85428 7.00592 3.90753 6.00884C3.97142 4.83339 4.31487 3.73115 5.04437 2.78467C5.93095 1.63318 7.15699 1.09005 8.56141 0.967577C10.5582 0.79319 12.555 0.982221 14.5518 0.927641C14.7102 0.927641 14.7462 0.99287 14.7449 1.13664C14.7449 2.581 14.7449 4.02668 14.7449 5.47104C14.7449 5.67604 14.6517 5.68669 14.4946 5.68669C13.4523 5.68669 12.4113 5.68669 11.3703 5.68669C10.3506 5.68669 9.92057 6.10868 9.90593 7.13904C9.89661 7.7647 9.91525 8.39303 9.89794 9.01869C9.88995 9.26364 9.96583 9.31822 10.2015 9.31688C11.7204 9.30623 13.2393 9.31688 14.7595 9.3049C15.0257 9.3049 15.0723 9.3728 15.0444 9.62439C14.89 10.9849 14.7515 12.3467 14.6144 13.7085C14.5691 14.1571 14.5785 14.1585 14.1458 14.1585C12.8386 14.1585 11.5313 14.1665 10.2254 14.1518C9.95119 14.1518 9.89794 14.2317 9.89794 14.4899C9.90593 19.0799 9.89794 23.6752 9.91125 28.2612C9.91125 28.5674 9.8407 28.646 9.53186 28.6433C7.77866 28.6273 6.02414 28.6366 4.27094 28.634C3.82499 28.634 3.87158 28.6992 3.87158 28.22C3.87602 25.9472 3.87913 23.6739 3.8809 21.4002Z"> </path> </svg> <span class="usa-sr-only">NCBI on Facebook</span> </a> <a href="https://www.linkedin.com/company/ncbinlm" class="ncbi-footer__social-icon ncbi-footer__social-icon--gray" target="_blank" rel="noreferrer noopener"> <svg width="25" height="23" viewBox="0 0 26 24" fill="none" xmlns="http://www.w3.org/2000/svg" focusable="false" aria-hidden="true"> <path d="M14.6983 9.98423C15.6302 9.24808 16.5926 8.74754 17.6762 8.51991C19.673 8.09126 21.554 8.30824 23.1262 9.7526C24.2351 10.7723 24.7529 12.1115 25.0165 13.5612C25.1486 14.3363 25.2105 15.1218 25.2015 15.9081C25.2015 18.3043 25.2015 20.6898 25.2082 23.0806C25.2082 23.3468 25.1549 23.444 24.8621 23.4414C23.1297 23.4272 21.3992 23.4272 19.6704 23.4414C19.4041 23.4414 19.3429 23.3588 19.3442 23.1019C19.3535 20.5194 19.3442 17.9368 19.3442 15.3543C19.3442 14.0005 18.3258 12.9448 17.0266 12.9488C15.7273 12.9528 14.6983 14.0071 14.6983 15.361C14.6983 17.9328 14.6917 20.5047 14.6983 23.0753C14.6983 23.3708 14.6198 23.444 14.3296 23.4427C12.6185 23.4294 10.9079 23.4294 9.19779 23.4427C8.93155 23.4427 8.86099 23.3735 8.86232 23.1086C8.8783 19.7619 8.88628 16.4144 8.88628 13.066C8.88628 11.5688 8.87874 10.0708 8.86365 8.57182C8.86365 8.3575 8.90758 8.27896 9.14054 8.28029C10.9048 8.29094 12.6687 8.29094 14.4321 8.28029C14.6464 8.28029 14.6983 8.34818 14.6983 8.54653C14.6903 9.00047 14.6983 9.45441 14.6983 9.98423Z"> </path> <path d="M6.55316 15.8443C6.55316 18.2564 6.55316 20.6699 6.55316 23.082C6.55316 23.3629 6.48127 23.4388 6.19906 23.4374C4.47737 23.4241 2.75568 23.4241 1.03399 23.4374C0.767751 23.4374 0.69986 23.3629 0.701191 23.1006C0.709178 18.2648 0.709178 13.4281 0.701191 8.59053C0.701191 8.34026 0.765089 8.27237 1.01669 8.2737C2.74991 8.28435 4.48048 8.28435 6.20838 8.2737C6.47462 8.2737 6.5465 8.33627 6.54517 8.6065C6.54783 11.0186 6.55316 13.4308 6.55316 15.8443Z"> </path> <path d="M3.65878 0.243898C5.36804 0.243898 6.58743 1.45529 6.58743 3.1406C6.58743 4.75801 5.32145 5.95742 3.60819 5.96807C3.22177 5.97614 2.83768 5.90639 2.47877 5.76299C2.11985 5.61959 1.79344 5.40546 1.51897 5.13334C1.24449 4.86123 1.02755 4.53668 0.881058 4.17902C0.734563 3.82136 0.661505 3.43788 0.666231 3.05141C0.67555 1.42601 1.9362 0.242566 3.65878 0.243898Z"> </path> </svg> <span class="usa-sr-only">NCBI on LinkedIn</span> </a> <a href="https://github.com/ncbi" class="ncbi-footer__social-icon ncbi-footer__social-icon--gray" target="_blank" rel="noreferrer noopener"> <svg width="28" height="27" viewBox="0 0 28 28" fill="none" xmlns="http://www.w3.org/2000/svg" focusable="false" aria-hidden="true"> <path d="M16.7228 20.6334C17.5057 20.5527 18.2786 20.3944 19.0301 20.1608C21.3108 19.4193 22.5822 17.8259 22.963 15.4909C23.1228 14.5112 23.1814 13.5287 22.9883 12.5437C22.8106 11.6423 22.4013 10.8028 21.8007 10.1076C21.7526 10.0605 21.7197 10 21.7064 9.934C21.6931 9.86799 21.7 9.79952 21.7262 9.73748C22.0856 8.6206 21.9711 7.51969 21.601 6.42677C21.582 6.3497 21.5345 6.2827 21.468 6.23923C21.4016 6.19577 21.3211 6.17906 21.2429 6.19248C20.7329 6.21649 20.2313 6.33051 19.7611 6.52928C19.1103 6.7908 18.4899 7.12198 17.9104 7.51703C17.84 7.56996 17.7581 7.60551 17.6713 7.62078C17.5846 7.63605 17.4954 7.6306 17.4112 7.60489C15.2596 7.05882 13.0054 7.06203 10.8554 7.61421C10.7806 7.63586 10.7018 7.63967 10.6253 7.62534C10.5487 7.611 10.4766 7.57892 10.4148 7.53167C9.64788 7.03247 8.85171 6.58918 7.96368 6.33359C7.65781 6.24338 7.34123 6.19458 7.02239 6.18849C6.94879 6.17986 6.87462 6.19893 6.81432 6.242C6.75402 6.28507 6.71191 6.34904 6.69621 6.42145C6.32342 7.51437 6.2209 8.61527 6.56307 9.73348C6.59635 9.84264 6.64694 9.93316 6.54177 10.0516C5.47666 11.2604 5.09988 12.6834 5.19574 14.2676C5.2663 15.4244 5.46201 16.5466 6.01454 17.5769C6.84399 19.1171 8.21664 19.9119 9.85158 20.3352C10.3938 20.4706 10.9444 20.5698 11.4998 20.632C11.5384 20.7492 11.4506 20.7798 11.408 20.8291C11.1734 21.1179 10.9894 21.4441 10.8634 21.7942C10.7622 22.0458 10.8315 22.4039 10.6065 22.5516C10.263 22.7766 9.83827 22.8485 9.42421 22.8871C8.17936 23.0056 7.26471 22.4877 6.6283 21.4348C6.25552 20.8184 5.76956 20.3325 5.08523 20.0663C4.76981 19.9325 4.42139 19.8967 4.08537 19.9638C3.7898 20.029 3.73788 20.1901 3.93891 20.4111C4.03639 20.5234 4.14989 20.6207 4.27575 20.6999C4.9796 21.1318 5.51717 21.7884 5.80152 22.5636C6.37002 23.9973 7.48039 24.5697 8.93825 24.6323C9.43741 24.6575 9.93768 24.615 10.4254 24.5058C10.5892 24.4672 10.6531 24.4872 10.6517 24.6762C10.6451 25.4936 10.6637 26.3123 10.6517 27.131C10.6517 27.6635 10.1684 27.9297 9.58663 27.7393C8.17396 27.2671 6.84977 26.5631 5.66838 25.656C2.59555 23.2891 0.720966 20.1861 0.217704 16.3376C-0.357453 11.9127 0.911353 8.00824 3.98551 4.73881C6.11909 2.42656 8.99932 0.939975 12.1203 0.540191C16.5351 -0.0601815 20.4347 1.14323 23.7232 4.16373C26.2449 6.47869 27.724 9.37672 28.1048 12.7726C28.5828 17.0325 27.3686 20.7945 24.4768 23.9827C22.9762 25.6323 21.0956 26.8908 18.9982 27.6488C18.8783 27.6927 18.7585 27.738 18.636 27.7726C18.0356 27.9404 17.6189 27.6395 17.6189 27.0098C17.6189 25.7452 17.6308 24.4806 17.6295 23.2159C17.6329 22.9506 17.6128 22.6856 17.5696 22.4238C17.4325 21.6664 17.3419 21.484 16.7228 20.6334Z"> </path> </svg> <span class="usa-sr-only">NCBI on GitHub</span> </a> <a href="https://ncbiinsights.ncbi.nlm.nih.gov/" class="ncbi-footer__social-icon ncbi-footer__social-icon--gray" target="_blank" rel="noreferrer noopener"> <svg width="26" height="26" viewBox="0 0 27 27" fill="none" xmlns="http://www.w3.org/2000/svg" focusable="false" aria-hidden="true"> <path d="M23.7778 26.4574C23.1354 26.3913 22.0856 26.8024 21.636 26.3087C21.212 25.8444 21.4359 24.8111 21.324 24.0347C19.9933 14.8323 14.8727 8.80132 6.09057 5.85008C4.37689 5.28406 2.58381 4.99533 0.779072 4.99481C0.202773 4.99481 -0.0229751 4.83146 0.00455514 4.21479C0.0660406 3.08627 0.0660406 1.95525 0.00455514 0.826734C-0.0413285 0.0815827 0.259669 -0.0193618 0.896534 0.00266238C6.96236 0.222904 12.3693 2.24179 16.9889 6.16209C22.9794 11.2478 26.1271 17.7688 26.4372 25.648C26.4629 26.294 26.3179 26.5271 25.6609 26.4684C25.0827 26.417 24.4991 26.4574 23.7778 26.4574Z"> </path> <path d="M14.8265 26.441C14.0924 26.441 13.2371 26.6795 12.6626 26.3786C12.0092 26.0372 12.3781 25.0644 12.246 24.378C11.1154 18.5324 6.6849 14.5497 0.74755 14.1001C0.217135 14.0615 -0.0104482 13.9422 0.0134113 13.3659C0.0519536 12.1454 0.0482829 10.9213 0.0134113 9.69524C-0.00127145 9.14464 0.196946 9.03268 0.703502 9.04736C9.21217 9.27128 16.5994 16.2511 17.2804 24.7231C17.418 26.4446 17.418 26.4446 15.6579 26.4446H14.832L14.8265 26.441Z"> </path> <path d="M3.58928 26.4555C2.64447 26.4618 1.73584 26.0925 1.06329 25.4289C0.39073 24.7653 0.00933763 23.8617 0.0030097 22.9169C-0.00331824 21.9721 0.365937 21.0635 1.02954 20.3909C1.69315 19.7184 2.59675 19.337 3.54156 19.3306C4.48637 19.3243 5.39499 19.6936 6.06755 20.3572C6.7401 21.0208 7.1215 21.9244 7.12782 22.8692C7.13415 23.814 6.7649 24.7226 6.10129 25.3952C5.43768 26.0677 4.53409 26.4491 3.58928 26.4555Z"> </path> </svg> <span class="usa-sr-only">NCBI RSS feed</span> </a> </div> </div> <div data-testid="gridContainer" class="grid-container ncbi-footer__container"> <div class="grid-row ncbi-footer__main-content-container" data-testid="grid"> <div class="ncbi-footer__column"> <p class="ncbi-footer__circled-icons-heading"> Connect with NLM </p> <div class="ncbi-footer__circled-icons-list"> <a href=https://twitter.com/nlm_nih class="ncbi-footer__social-icon ncbi-footer__social-icon--circled" target="_blank" rel="noreferrer noopener"> <svg width="32" height="32" viewBox="0 0 40 40" fill="none" xmlns="http://www.w3.org/2000/svg" focusable="false" aria-hidden="true"> <path d="m6.067 8 10.81 13.9L6 33.2h4.2l8.4-9.1 7.068 9.1H34L22.8 18.5 31.9 8h-3.5l-7.7 8.4L14.401 8H6.067Zm3.6 1.734h3.266l16.8 21.732H26.57L9.668 9.734Z"> </path> </svg> <span class="usa-sr-only">NLM on X (formerly known as Twitter)</span> </a> <a href=https://www.facebook.com/nationallibraryofmedicine class="ncbi-footer__social-icon ncbi-footer__social-icon--circled" target="_blank" rel="noreferrer noopener"> <svg width="13" height="24" viewBox="0 0 13 24" fill="none" xmlns="http://www.w3.org/2000/svg" focusable="false" aria-hidden="true"> <path d="M4.11371 23.1369C4.11371 23.082 4.11371 23.0294 4.11371 22.9745V12.9411H0.817305C0.6709 12.9411 0.670898 12.9411 0.670898 12.8016C0.670898 11.564 0.670898 10.3287 0.670898 9.09341C0.670898 8.97903 0.705213 8.95158 0.815017 8.95158C1.8673 8.95158 2.91959 8.95158 3.97417 8.95158H4.12057V8.83263C4.12057 7.8055 4.12057 6.7738 4.12057 5.74897C4.1264 4.92595 4.31387 4.11437 4.66959 3.37217C5.12916 2.38246 5.94651 1.60353 6.95717 1.1921C7.64827 0.905008 8.3913 0.764035 9.13953 0.778051C10.0019 0.791777 10.8644 0.830666 11.7268 0.860404C11.8869 0.860404 12.047 0.894717 12.2072 0.90158C12.2964 0.90158 12.3261 0.940469 12.3261 1.02968C12.3261 1.5421 12.3261 2.05452 12.3261 2.56465C12.3261 3.16857 12.3261 3.7725 12.3261 4.37642C12.3261 4.48165 12.2964 4.51367 12.1912 4.51138C11.5369 4.51138 10.8804 4.51138 10.2261 4.51138C9.92772 4.51814 9.63058 4.5526 9.33855 4.61433C9.08125 4.6617 8.84537 4.78881 8.66431 4.97766C8.48326 5.16652 8.3662 5.40755 8.32972 5.66661C8.28476 5.89271 8.26027 6.1224 8.25652 6.35289C8.25652 7.19014 8.25652 8.02969 8.25652 8.86923C8.25652 8.89439 8.25652 8.91955 8.25652 8.95615H12.0219C12.1797 8.95615 12.182 8.95616 12.1614 9.10714C12.0768 9.76596 11.9876 10.4248 11.9029 11.0813C11.8312 11.6319 11.7626 12.1824 11.697 12.733C11.6719 12.9434 11.6787 12.9434 11.4683 12.9434H8.26338V22.899C8.26338 22.979 8.26338 23.0591 8.26338 23.1392L4.11371 23.1369Z"> </path> </svg> <span class="usa-sr-only">NLM on Facebook</span> </a> <a href=https://www.youtube.com/user/NLMNIH class="ncbi-footer__social-icon ncbi-footer__social-icon--circled" target="_blank" rel="noreferrer noopener"> <svg width="21" height="15" viewBox="0 0 21 15" fill="none" xmlns="http://www.w3.org/2000/svg" focusable="false" aria-hidden="true"> <path d="M19.2561 1.47914C18.9016 1.15888 18.5699 0.957569 17.2271 0.834039C15.5503 0.678484 13.2787 0.655608 11.563 0.65332H9.43556C7.71987 0.65332 5.4483 0.678484 3.77151 0.834039C2.43098 0.957569 2.097 1.15888 1.74242 1.47914C0.813665 2.32097 0.619221 4.62685 0.598633 6.89384C0.598633 7.31781 0.598633 7.74101 0.598633 8.16345C0.626084 10.4121 0.827391 12.686 1.74242 13.521C2.097 13.8412 2.4287 14.0425 3.77151 14.1661C5.4483 14.3216 7.71987 14.3445 9.43556 14.3468H11.563C13.2787 14.3468 15.5503 14.3216 17.2271 14.1661C18.5676 14.0425 18.9016 13.8412 19.2561 13.521C20.1712 12.6929 20.3725 10.451 20.3999 8.22064C20.3999 7.74025 20.3999 7.25986 20.3999 6.77946C20.3725 4.54907 20.1689 2.30724 19.2561 1.47914ZM8.55942 10.5311V4.65201L13.5601 7.50005L8.55942 10.5311Z" fill="white" /> </svg> <span class="usa-sr-only">NLM on YouTube</span> </a> </div> </div> <address class="ncbi-footer__address ncbi-footer__column"> <p> <a class="usa-link usa-link--external" href="https://www.google.com/maps/place/8600+Rockville+Pike,+Bethesda,+MD+20894/%4038.9959508, -77.101021,17z/data%3D!3m1!4b1!4m5!3m4!1s0x89b7c95e25765ddb%3A0x19156f88b27635b8!8m2!3d38.9959508! 4d-77.0988323" rel="noopener noreferrer" target="_blank">National Library of Medicine <br/> 8600 Rockville Pike<br/> Bethesda, MD 20894</a> </p> </address> <ul class="usa-list usa-list--unstyled ncbi-footer__vertical-list ncbi-footer__column"> <li class="ncbi-footer__vertical-list-item"> <a href="https://www.nlm.nih.gov/web_policies.html" class="usa-link usa-link--alt ncbi-footer__link" > Web Policies </a> </li> <li class="ncbi-footer__vertical-list-item"> <a href="https://www.nih.gov/institutes-nih/nih-office-director/office-communications-public-liaison/freedom-information-act-office" class="usa-link usa-link--alt ncbi-footer__link" > FOIA </a> </li> <li class="ncbi-footer__vertical-list-item"> <a href="https://www.hhs.gov/vulnerability-disclosure-policy/index.html" class="usa-link usa-link--external usa-link--alt ncbi-footer__link" rel="noreferrer noopener" target='_blank' > HHS Vulnerability Disclosure </a> </li> </ul> <ul class="usa-list usa-list--unstyled ncbi-footer__vertical-list ncbi-footer__column"> <li class="ncbi-footer__vertical-list-item"> <a href="https://support.nlm.nih.gov/" class="usa-link usa-link--alt ncbi-footer__link" > Help </a> </li> <li class="ncbi-footer__vertical-list-item"> <a href="https://www.nlm.nih.gov/accessibility.html" class="usa-link usa-link--alt ncbi-footer__link" > Accessibility </a> </li> <li class="ncbi-footer__vertical-list-item"> <a href="https://www.nlm.nih.gov/careers/careers.html" class="usa-link usa-link--alt ncbi-footer__link" > Careers </a> </li> </ul> </div> <div class="grid-row grid-col-12" data-testid="grid"> <ul class="ncbi-footer__bottom-links-list"> <li class="ncbi-footer__bottom-list-item"> <a href="https://www.nlm.nih.gov/" class="usa-link usa-link--alt ncbi-footer__link" > NLM </a> </li> <li class="ncbi-footer__bottom-list-item"> <a href="https://www.nih.gov/" class="usa-link usa-link--alt ncbi-footer__link" > NIH </a> </li> <li class="ncbi-footer__bottom-list-item"> <a href="https://www.hhs.gov/" class="usa-link usa-link--external usa-link--alt ncbi-footer__link" rel="noreferrer noopener" target='_blank' > HHS </a> </li> <li class="ncbi-footer__bottom-list-item"> <a href="https://www.usa.gov/" class="usa-link usa-link--external usa-link--alt ncbi-footer__link" rel="noreferrer noopener" target='_blank' > USA.gov </a> </li> </ul> </div> </div> </footer> <script type="text/javascript" src="https://cdn.ncbi.nlm.nih.gov/core/pinger/pinger.js"> </script> <button class="back-to-top" data-ga-category="pagination" data-ga-action="back_to_top"> <label>Back to Top</label> <svg class="usa-icon order-0" aria-hidden="true" focusable="false" role="img"> <use xlink:href="/static/img/sprite.svg#arrow_upward"></use> </svg> </button> <script src="https://code.jquery.com/jquery-3.5.0.min.js" integrity="sha256-xNzN2a4ltkB44Mc/Jz3pT4iU1cmeR0FkXs4pru/JxaQ=" crossorigin="anonymous"> </script> <script type="text/javascript">var exports = {};</script> <script src="/static/CACHE/js/output.13b077bc3ffd.js"></script> <script type="application/javascript"> window.ncbi = window.ncbi || {}; window.ncbi.pmc = window.ncbi.pmc || {}; window.ncbi.pmc.options = { logLevel: 'INFO', staticEndpoint: '/static/', citeCookieName: 'pmc-cf', }; </script> <script type="module" crossorigin="" src="/static/assets/base-9bea7450.js"></script> <script type="module" crossorigin="" src="/static/assets/article-722d91a2.js"></script> <script type="module" crossorigin="" src="/static/assets/math-3a787fba.js"></script> </body> </html>

CINXE.COM

A high-content image-based drug screen of clinical compounds against cell transmission of adenovirus - PMC