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<select id="email-citation-format" name="citation-format" class="action-panel-selector email-citation-format">
<option selected="selected" value="summary">Summary</option>
<option value="summary-text">Summary (text)</option>
<option value="abstract">Abstract</option>
<option value="abstract-text">Abstract (text)</option>
</select>
</div>
<div class="include-supplemental-container">
<input type="checkbox" aria-label="Include MeSH and other data" name="include-supplemental" id="email-include-supplemental" class="email-include-supplemental">
<label for="email-include-supplemental" class="email-include-supplemental-label">MeSH and other data</label>
</div>
<div class="form-field recaptcha ">
<div class="g-recaptcha" id="id-recaptcha" data-sitekey="6LfsWHMdAAAAAClKbtOpjQ2pMjgsGxvv7NdZW9uI"></div>
</div>
<div id="captcha-error-message" class="usa-input-error-message captcha-validation-message" role="alert"></div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="send">
Send email
</button>
<button class="action-panel-cancel"
aria-label="Close 'Email citations' panel"
ref="linksrc=close_email_panel"
aria-controls="email-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="cancel">
Cancel
</button>
</div>
<input type="hidden" name="email-search-details" value="" />
<input type="hidden" name="email-search-details-hash" value="0e42663a6c3bd85498fcb88798998fed7bfdc45d457db35281e41afe13cc0524" />
</form>
</div>
</div>
<div id="collections-action-panel"
class="collections-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-collections-open-panel-enabled="false"
data-collections-open-panel-url-hash="#open-collections-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to Collections
</h3>
<form id="collections-action-panel-form"
class="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-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-myncbi-max-collection-name-length="100"
data-add-to-collection-max-amount="1000"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/">
<input type="hidden" name="csrfmiddlewaretoken" value="mTG052OCphuK6SmnxAYnz95wfqJv2hoJDGq5IShYZShe0jsdaTBp3hBaNurHVSW3">
<div class="choice-group" role="radiogroup">
<ul class="radio-group-items">
<li>
<input type="radio"
id="collections-action-panel-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_new">
<label for="collections-action-panel-new">Create a new collection</label>
</li>
<li>
<input type="radio"
id="collections-action-panel-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_existing">
<label for="collections-action-panel-existing">Add to an existing collection</label>
</li>
</ul>
</div>
<div class="controls-wrapper">
<div class="action-panel-control-wrap new-collections-controls">
<label for="collections-action-panel-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label>
<input
type="text"
name="add-to-new-collection"
id="collections-action-panel-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="save_share"
data-ga-action="create_collection"
data-ga-label="non_favorties_collection">
<div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
Name must be less than 100 characters
</div>
</div>
<div class="action-panel-control-wrap existing-collections-controls">
<label for="collections-action-panel-add-to-existing" class="action-panel-label">
Choose a collection:
</label>
<select id="collections-action-panel-add-to-existing"
class="action-panel-selector collections-action-add-to-existing"
data-ga-category="save_share"
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>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="add">
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to Collections' panel"
ref="linksrc=close_collections_panel"
aria-controls="collections-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="bibliography-action-panel"
class="bibliography-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-bibliography-open-panel-enabled="false"
data-bibliography-open-panel-url-hash="#open-bibliography-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to My Bibliography
</h3>
<form id="bibliography-action-panel-form"
class="bibliography-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-add-to-bibliography-max-amount="100"
data-add-to-bibliography-batch-size="10"
data-bibliography-delegates-url="/list-bibliography-delegates/"
data-add-to-bibliography-url="/add-to-bibliography/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-mybib-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography/">
<input type="hidden" name="csrfmiddlewaretoken" value="mTG052OCphuK6SmnxAYnz95wfqJv2hoJDGq5IShYZShe0jsdaTBp3hBaNurHVSW3">
<div class="action-panel-control-wrap bibliographies-controls">
<div class="choice-group">
<ul class="bibliographies-action-add radio-group-items">
<li>
<input name="bibliography" id="my-bibliography" class="my-bibliography" type="radio" checked/>
<label for="my-bibliography">My Bibliography</label>
</li>
</ul>
</div>
</div>
<div class="bibliographies-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your delegates due to an error<br>
<a href="#">Please try again</a>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore>
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to bibliography' panel"
ref="linksrc=close_bibliography_panel"
aria-controls="bibliography-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="mybib"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="saved-search-action-panel" class="saved-search-action-panel action-panel " aria-hidden="true"
data-saved-search-open-panel-enabled="false"
data-saved-search-open-panel-url-hash="#open-saved-search-panel">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your saved search
</h2>
<form id="saved-search-action-panel-form"
class="saved-search-action-panel-form action-panel-content action-form"
data-create-saved-search-url="/create-saved-search/"
data-try-search-terms-url="/try-search-term/"
data-saved-search-root-url="https://www.ncbi.nlm.nih.gov/myncbi/searches/">
<input type="hidden" name="csrfmiddlewaretoken" value="mTG052OCphuK6SmnxAYnz95wfqJv2hoJDGq5IShYZShe0jsdaTBp3hBaNurHVSW3">
<div class="action-panel-control-wrap">
<label for="saved-search-name" class="action-panel-label saved-search-name-label required-field-asterisk">
Name of saved search:
</label>
<input maxlength="200"
type="text"
name="saved-search-name"
id="saved-search-name"
class="saved-search-name"
value=""
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-term" class="action-panel-label required-field-asterisk">
Search terms:
</label>
<textarea name="saved-search-term" id="saved-search-term" class="saved-search-term" required></textarea>
</div>
<div class="test-search-term-wrap">
<a href="#" class="try-search-term">Test search terms</a>
</div>
<div class="choice-group action-panel-extra-margin-top">
<span class="action-panel-label" id="fieldset-label">
Would you like email updates of new search results?
</span>
<fieldset id="saved-search-alert" aria-describedby="fieldset-label">
<legend class="usa-sr-only">Saved Search Alert Radio Buttons</legend>
<ul class="radio-group-items">
<li>
<input type="radio" id="saved-search-alert-yes" class="saved-search-alert-yes" name="saved-search-alert" value="yes" checked>
<label for="saved-search-alert-yes" class="action-panel-label">Yes</label>
</li>
<li>
<input aria-label="No radio input" type="radio" id="saved-search-alert-no" class="saved-search-alert-no" name="saved-search-alert" value="no">
<label for="saved-search-alert-no" class="action-panel-label">No</label>
</li>
</ul>
</fieldset>
</div>
<div class="alert-schedule-wrap">
<div class="action-panel-control-wrap">
<label class="action-panel-label">
Email:
</label>
<span aria-label="Email address" id="saved-search-email" class="action-panel-label"><span class="action-panel-label-bold"></span> (<a class="myncbi-account-settings" href="https://www.ncbi.nlm.nih.gov/account/settings/">change</a>)</span>
</div>
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="saved-search-frequency" class="action-panel-label">
Frequency:
</label>
<select id="saved-search-frequency" class="no-border-panel-selector saved-search-frequency">
<option value="monthly">Monthly</option>
<option value="weekly">Weekly</option>
<option value="daily">Daily</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-monthly-additional">
<label for="saved-search-monthly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-monthly-on-day" class="no-border-panel-selector">
<option value="Sunday">The first Sunday</option>
<option value="Monday">The first Monday</option>
<option value="Tuesday">The first Tuesday</option>
<option value="Wednesday">The first Wednesday</option>
<option value="Thursday">The first Thursday</option>
<option value="Friday">The first Friday</option>
<option value="Saturday">The first Saturday</option>
<option value="day">The first day</option>
<option value="weekday">The first weekday</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-weekly-additional">
<label for="saved-search-weekly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-weekly-on-day" class="no-border-panel-selector saved-search-weekly-on-day">
<option value="Sunday">Sunday</option>
<option value="Monday">Monday</option>
<option value="Tuesday">Tuesday</option>
<option value="Wednesday">Wednesday</option>
<option value="Thursday">Thursday</option>
<option value="Friday">Friday</option>
<option value="Saturday">Saturday</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-report" class="action-panel-label">
Report format:
</label>
<select id="saved-search-report" class="no-border-panel-selector saved-search-report">
<option value="DocSum">Summary</option>
<option value="DocSumText">Summary (text)</option>
<option value="Abstract">Abstract</option>
<option value="AbstractText">Abstract (text)</option>
<option value="MEDLINE">PubMed</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-amount" class="action-panel-label">
Send at most:
</label>
<select id="saved-search-amount" class="no-border-panel-selector saved-search-amount">
<option value="1">1 item</option>
<option value="5" selected>5 items</option>
<option value="10">10 items</option>
<option value="20">20 items</option>
<option value="50">50 items</option>
<option value="100">100 items</option>
<option value="200">200 items</option>
</select>
</div>
<div>
<input type="checkbox" id="saved-search-send-if-no-result" class="saved-search-send-if-no-result" name="saved-search-send-if-no-result">
<label for="saved-search-send-if-no-result" class="action-panel-label smaller-checkbox">
Send even when there aren't any new results
</label>
</div>
<div class="action-panel-control-wrap option-text-in-email-wrap">
<label for="saved-search-email-text" class="action-panel-label">
Optional text in email:
</label>
<textarea name="saved-search-email-text"
id="saved-search-email-text"
class="saved-search-email-text"></textarea>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Saving..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Save
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your saved search' panel"
ref="linksrc=close_saved_search_panel"
aria-controls="saved-search-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="citation-manager-action-panel" class="citation-manager-action-panel action-panel" aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Create a file for external citation management software
</h2>
<form id="citation-manager-action-panel-form"
class="action-panel-content action-form"
action="/results-export-ids/"
data-by-search-action="/results-export-search-data/"
data-by-ids-action="/results-export-ids/"
method="post"
data-by-search-method="post"
data-by-ids-method="post">
<input type="hidden" name="csrfmiddlewaretoken" value="mTG052OCphuK6SmnxAYnz95wfqJv2hoJDGq5IShYZShe0jsdaTBp3hBaNurHVSW3">
<input name="results-format" type="hidden" value="pubmed"/>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="save">
Create file
</button>
<button class="action-panel-cancel"
aria-label="Close 'Send citations to citation manager' panel"
ref="linksrc=close_citation_manager_panel"
aria-controls="citation-manager-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="rss-action-panel" class="rss-action-panel action-panel " aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your RSS Feed
</h2>
<form id="rss-action-panel-form"
class="rss-action-panel-form action-panel-content action-form"
data-create-rss-feed-url="/create-rss-feed-url/"
data-search-form-term-value="">
<input type="hidden" name="csrfmiddlewaretoken" value="mTG052OCphuK6SmnxAYnz95wfqJv2hoJDGq5IShYZShe0jsdaTBp3hBaNurHVSW3">
<div class="action-panel-control-wrap">
<label for="rss-name" class="action-panel-label required-field-asterisk">
Name of RSS Feed:
</label>
<input maxlength="200"
placeholder="Name"
type="text"
name="rss-name"
id="rss-name"
class="rss-name"
value=''
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="rss-limit-wrap">
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="rss-limit" class="action-panel-label">
Number of items displayed:
</label>
<select id="rss-limit" class="no-border-panel-selector rss-limit">
<option value="5">5</option>
<option value="10">10</option>
<option value="15" selected="selected">15</option>
<option value="20">20</option>
<option value="50">50</option>
<option value="100">100</option>
</select>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Creating..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Create RSS
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your RSS' panel"
ref="linksrc=close_rss_panel"
aria-controls="rss-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
<div class="action-panel-control-wrap rss-link-copy-wrap">
<label for="rss-link" class="usa-sr-only">RSS Link</label>
<input placeholder="Your RSS Feed Link" type="text" name="rss-link" id="rss-link" class="rss-link" title="RSS Link">
<button
type="button"
disabled
class="rss-link-copy-button disabled"
data-ga-category="save_share"
data-ga-action="rss"
data-ga-label="copy">
Copy
</button>
</div>
</form>
</div>
</div>
</div>
</div>
<div class="article-page" id="article-page" data-article-pmid="31263175">
<aside class="page-sidebar">
<div class="inner-wrap">
<div class="full-text-links">
<div class="full-view">
<h3 class="title">
Full text links
</h3>
<div class="full-text-links-list">
<a class="link-item
dialog-focus"
href="https://doi.org/10.1038/s41418-019-0378-6"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=3094&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=31263175&amp;db=pubmed&amp;nlmid=9437445"
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data-ga-action="Nature Publishing Group"
data-ga-label="31263175"
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Nature Publishing Group
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/31263175/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=31263175&amp;db=pubmed&amp;nlmid=9437445"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="31263175"
><span class="text">
Free PMC article
</span></a>
</div>
</div>
<div class="short-view">
<a href="#" class="full-text-links-button full-text-links-dialog-trigger">
Full text links
</a>
</div>
</div>
<div class="actions-buttons sidebar"><h3 class="title">Actions</h3><div class="inner-wrap"><button class="citation-button citation-dialog-trigger"
aria-label="Open dialog with citation text in different styles"
data-ga-category="save_share"
data-ga-action="cite"
data-ga-label="open"
data-all-citations-url="/31263175/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/31263175/export/"><span class="button-label">Cite</span></button><link type="text/css" href="ncbi-overlay-block/src/overlay-block.css"><div class="collections-button-container" data-article-id="31263175" data-article-db="pubmed"><button class="collections-button collections-dialog-trigger"
aria-label="Save article in MyNCBI collections."
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_button"
data-collections-open-dialog-enabled="false"
data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F31263175%2F%23open-collections-dialog"
data-in-collections="false"><span class="button-label">Collections</span></button><div class="overlay" role="dialog"><div id="collections-action-dialog"
class="dialog collections-dialog"
aria-hidden="true"><div class="title">Add to Collections</div><div class="collections-action-panel action-panel"><form id="collections-action-dialog-form"
class="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="mTG052OCphuK6SmnxAYnz95wfqJv2hoJDGq5IShYZShe0jsdaTBp3hBaNurHVSW3"><div class="choice-group" role="radiogroup"><ul class="radio-group-items"><li><input type="radio"
id="collections-action-dialog-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_new"><label for="collections-action-dialog-new">Create a new collection</label></li><li><input type="radio"
id="collections-action-dialog-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_existing"><label for="collections-action-dialog-existing">Add to an existing collection</label></li></ul></div><div class="controls-wrapper"><div class="action-panel-control-wrap new-collections-controls"><label for="collections-action-dialog-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label><input
type="text"
name="add-to-new-collection"
id="collections-action-dialog-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="collections_button"
data-ga-action="create_collection"
data-ga-label="non_favorties_collection"><div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
Name must be less than 100 characters
</div></div><div class="action-panel-control-wrap existing-collections-controls"><label for="collections-action-dialog-add-to-existing" class="action-panel-label">
Choose a collection:
</label><select id="collections-action-dialog-add-to-existing"
class="action-panel-selector collections-action-add-to-existing"
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<h1 class="heading-title">
ER complex proteins are required for rhodopsin biosynthesis and photoreceptor survival in Drosophila and mice
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</a><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, China. xjzhu@uestc.edu.cn." href="#short-view-affiliation-8" ref="linksrc=author_aff">
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</a><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Department of Ophthalmology, First People&#x27;s Hospital of Shangqiu, Shangqiu, Henan, China. xjzhu@uestc.edu.cn." href="#short-view-affiliation-10" ref="linksrc=author_aff">
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><sup class="key">3</sup> Sichuan Provincial Key Laboratory for Human Disease Gene Study, Sichuan Provincial People&#x27;s Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, 610072, China.</li>
<li data-affiliation-id="short-view-affiliation-4"
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><sup class="key">4</sup> Southwest Jiaotong University, Chengdu, Sichuan, China.</li>
<li data-affiliation-id="short-view-affiliation-5"
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><sup class="key">5</sup> CAS Key Laboratory of Standardization and Measurement for Nanotechnology, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology, Beijing, 100190, China.</li>
<li data-affiliation-id="short-view-affiliation-6"
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><sup class="key">6</sup> Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua University, Beijing, 100871, China.</li>
<li data-affiliation-id="short-view-affiliation-7"
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><sup class="key">7</sup> Sichuan Provincial Key Laboratory for Human Disease Gene Study, Sichuan Provincial People&#x27;s Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, 610072, China. xjzhu@uestc.edu.cn.</li>
<li data-affiliation-id="short-view-affiliation-8"
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><sup class="key">8</sup> Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, China. xjzhu@uestc.edu.cn.</li>
<li data-affiliation-id="short-view-affiliation-9"
id="short-view-affiliation-9"
><sup class="key">9</sup> Chinese Academy of Sciences Sichuan Translational Medicine Hospital, Chengdu, China. xjzhu@uestc.edu.cn.</li>
<li data-affiliation-id="short-view-affiliation-10"
id="short-view-affiliation-10"
><sup class="key">10</sup> Department of Ophthalmology, First People&#x27;s Hospital of Shangqiu, Shangqiu, Henan, China. xjzhu@uestc.edu.cn.</li>
<li data-affiliation-id="short-view-affiliation-11"
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><sup class="key">11</sup> National Institute of Biological Sciences, Beijing, 102206, China. wangtao1006@nibs.ac.cn.</li>
<li data-affiliation-id="short-view-affiliation-12"
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><sup class="key">12</sup> Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua University, Beijing, 100871, China. wangtao1006@nibs.ac.cn.</li>
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Defective rhodopsin homeostasis is one of the major causes of retinal degeneration, including the disease Retinitis pigmentosa. To identify cellular factors required for the biosynthesis of rhodopsin, we performed a genome-wide genetic screen in Drosophila for mutants with reduced levels of rhodopsin. We isolated loss-of-function alleles in endoplasmic reticulum membrane protein complex 3 (emc3), emc5, and emc6, each of which exhibited defective phototransduction and photoreceptor cell degeneration. EMC3, EMC5, and EMC6 were essential for rhodopsin synthesis independent of the ER associated degradation (ERAD) pathway, which eliminates misfolded proteins. We generated null mutations for all EMC subunits, and further demonstrated that different EMC subunits play roles in different cellular functions. Conditional knockout of the Emc3 gene in mice led to mislocalization of rhodopsin protein and death of cone and rod photoreceptor cells. These data indicate conserved roles for EMC subunits in maintaining rhodopsin homeostasis and photoreceptor function, and suggest that retinal degeneration may also be caused by defects in early biosynthesis of rhodopsin.
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<div class="figure-caption-contents"><p> Rhodopsin biosynthesis defects in the… </p></div>
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<div class="figure-caption-contents"><p> Rhodopsin biosynthesis defects in the <i> emc3 </i> , <i> emc5 </i> , and <i> emc6 </i> mutants. <b> a… </b> </p></div>
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<div class="figure-caption-contents">Rhodopsin biosynthesis defects in the <i>emc3</i>, <i>emc5</i>, and <i>emc6</i> mutants. <b>a</b> Isolation of the <i>emc3</i><sup><i>G7</i></sup>, <i>emc3</i><sup><i>G10</i></sup>, <i>emc5</i><sup><i>G2</i></sup>, and <i>emc6</i><sup><i>N10</i></sup> mutations via a forward genetic screen. Rh1-GFP fluorescence in the deep pseudopupil of 1-day-old wild-type (<i>ey-flp rh1-GFP;FRT40A/GMR-hid CL FRT40A</i>), <i>emc3</i><sup><i>G7</i></sup> (<i>ey-flp rh1-GFP;emc3</i><sup><i>G7</i></sup>
<i>FRT40A/GMR-hid CL FRT40A</i>), <i>emc3</i><sup><i>G10</i></sup> (<i>ey-flp rh1-GFP;emc3</i><sup><i>G10</i></sup>
<i>FRT40A/GMR-hid CL FRT40A</i>), <i>emc5</i><sup><i>G2</i></sup> (<i>ey-flp rh1-GFP;emc5</i><sup><i>G2</i></sup>
<i>FRT40A/GMR-hid CL FRT40A</i>), and <i>emc6</i><sup><i>N10</i></sup> (<i>ey-flp rh1-GFP; FRT82B emc6</i><sup><i>N10</i></sup><i>/FRT82B GMR-hid CL</i>) flies are shown. Scale bar is 100μm. <b>b</b> The <i>emc3</i>, <i>emc5</i>, and <i>emc6</i> loci and mutations associated with the <i>emc3</i><sup><i>G7</i></sup>, <i>emc3</i><sup><i>G10</i></sup>, <i>emc5</i><sup><i>G2</i></sup>, and <i>emc6</i><sup><i>N10</i></sup> alleles. <b>c</b> Rhodopsin levels were reduced in <i>emc3</i><sup><i>G7</i></sup>, <i>emc3</i><sup><i>G10</i></sup>, <i>emc5</i><sup><i>G2</i></sup>, and <i>emc6</i><sup><i>N10</i></sup> flies, and were restored by <i>ninaE-emc3</i>, <i>ninaE-emc5</i>, and <i>ninaE-emc6</i> transgenes, respectively. Protein extracts from 1/4 head of each genotype (1 d after eclosion) were loaded and probed with antibodies against Rh1, ß-actin, TRP, and INAD</div>
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<div class="figure-caption-contents"><p> Loss of PDA in <i> emc3… </i> </p></div>
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<p> Fig. 2 </p>
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<div class="figure-caption-contents"><p> Loss of PDA in <i> emc3 </i> , <i> emc5 </i> , and <i> emc6 </i> mutants. <b> a </b> ERG… </p></div>
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Fig. 2
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<div class="figure-caption-contents">Loss of PDA in <i>emc3</i>, <i>emc5</i>, and <i>emc6</i> mutants. <b>a</b> ERG recordings in wild-type flies (<i>ey-flp rh1-GFP;FRT40A/GMR-hid CL FRT40A</i>) showed that PDA was induced by blue light and terminated by orange light (arrows). <b>b</b> PDA was eliminated in <i>ninaE</i><sup><i>P332</i></sup> flies and in <b>c</b>
<i>emc3</i><sup><i>G7</i></sup>, <b>e</b>
<i>emc3</i><sup><i>G10</i></sup>, <b>g</b>
<i>emc5</i><sup><i>G2</i></sup>, and <b>i</b>
<i>emc6</i><sup><i>N10</i></sup> flies. PDA was restored in (<b>d</b>) <i>emc3</i><sup><i>G7</i></sup><i>;ninaE-emc3</i>, <b>f</b>
<i>emc3</i><sup><i>G10</i></sup><i>;ninaE-emc3</i>, <b>h</b>
<i>emc5</i><sup><i>G2</i></sup><i>;ninaE-emc5</i>, and <b>j</b>
<i>ninaE-emc6;emc6</i><sup><i>N10</i></sup> flies. Flies ~1day after eclosion were dark-adapted for 2min and subsequently exposed to 5-s pulses of orange (<b>O</b>) or blue (<b>B</b>) light. At least 10 flies of each genotype were tested</div>
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alt="Fig. 3" />
</a>
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<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="804">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Fig. 3 </p>
</strong>
<div class="figure-caption-contents"><p> EMC3, EMC5, and EMC6 were… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 3 </p>
</strong>
<div class="figure-caption-contents"><p> EMC3, EMC5, and EMC6 were not required for maturation of the Rh1 protein.… </p></div>
</div>
<figcaption id="figure-caption-2" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 3
</strong>
<div class="figure-caption-contents">EMC3, EMC5, and EMC6 were not required for maturation of the Rh1 protein. <b>a</b> Rh1 localized to the rhabdomere region in <i>emc3</i>, <i>emc5</i>, and <i>emc6</i> mutant photoreceptor cells. Tangential resin-embedded retina sections of compound eyes from ~1-day-old wt, <i>emc3</i><sup><i>G7</i></sup>, <i>emc3</i><sup><i>G7</i></sup><i>;ninaE-emc3</i>, <i>emc5</i><sup><i>G2</i></sup>, <i>emc5</i><sup><i>G2</i></sup><i>;ninaE-emc5, emc6</i><sup><i>N10</i></sup>, and <i>ninaE-emc6;emc6</i><sup><i>N10</i></sup> flies were labeled using antibodies against Rh1. Rhabdomeres are indicated by arrows. Scale bar is 2μm. Illustration of photoreceptor cells within a single ommatidium is on the top right corner, and rhabdomeres of R1-R7 photoreceptor cells are indicated by numbers. <b>b</b> Deglycosylation of Rh1 is normal in <i>emc3</i>, <i>emc5</i>, and <i>emc6</i> mutants. Head extracts prepared from 1-day-old wild-type, <i>ninaA</i>, <i>dmppe</i><sup><i>e02905</i></sup>, <i>emc3</i><sup><i>G7</i></sup>, <i>emc5</i><sup><i>G2</i></sup>, and <i>emc6</i><sup><i>N10</i></sup> flies were probed with antibodies against Rh1, and mature and immature bands are indicated. Extracts from <i>ninaA</i> and <i>emc6</i><sup><i>N10</i></sup> flies (10 heads), <i>emc3</i><sup><i>G7</i></sup> and <i>emc5</i><sup><i>G2</i></sup> flies (5 heads), and wild-type and <i>dmppe</i><sup><i>e02905</i></sup> flies (0.25 of a head) were loaded. <b>c</b><b>e</b> Similar Rh1 levels were seen in (<b>c</b>) <i>emc3</i><sup><i>G7</i></sup> and <i>emc3</i><sup><i>G7</i></sup><i>;hrd1</i><sup><i>1</i></sup> flies (<i>ey-flp rh1-GFP;emc3</i><sup><i>G7</i></sup>
<i>FRT40A/GMR-hid CL FRT40A;FRT82B hrd1</i><sup><i>1</i></sup><i>/FRT82B GMR-hid CL</i>), <b>d</b>
<i>emc5</i><sup><i>G2</i></sup> and <i>emc5</i><sup><i>G2</i></sup><i>;hrd1</i><sup><i>1</i></sup> flies (<i>ey-flp rh1-GFP;emc5</i><sup><i>G2</i></sup>
<i>FRT40A/GMR-hid CL FRT40A;FRT82B hrd1</i><sup><i>1</i></sup><i>/FRT82B GMR-hid CL</i>), and <b>e</b>
<i>emc6</i><sup><i>N10</i></sup> and <i>emc6</i><sup><i>N10</i></sup>
<i>hrd1</i><sup><i>1</i></sup> (<i>ey-flp rh1-GFP; FRT82B emc6</i><sup><i>N10</i></sup>
<i>hrd1</i><sup><i>1</i></sup><i>/FRT82B GMR-hid CL</i>) flies. Protein extracts from 0.25 of a head from ~1-day-old flies were loaded</div>
</figcaption>
</figure>
<figure class="figure-item "
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itemprop="associatedMedia"
data-slide-index="3"
data-label-slug="fig-4">
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itemprop="contentUrl"
aria-describedby="figure-caption-3"
role="button"
data-image-width="788"
data-image-height="823"
data-image-alt="Fig. 4"
data-pmc-id="PMC7206144"
data-figure-id="Fig4">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-3"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e2a/7206144/edf823bb952c/41418_2019_378_Fig4_HTML.gif"
alt="Fig. 4" />
</a>
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<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="823">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Fig. 4 </p>
</strong>
<div class="figure-caption-contents"><p> Interaction among EMC subunits. <b> a… </b> </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 4 </p>
</strong>
<div class="figure-caption-contents"><p> Interaction among EMC subunits. <b> a </b> S2 cells were transiently transfected with <i> emc3-RFP </i> ,… </p></div>
</div>
<figcaption id="figure-caption-3" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 4
</strong>
<div class="figure-caption-contents">Interaction among EMC subunits. <b>a</b> S2 cells were transiently transfected with <i>emc3-RFP</i>, <i>emc5-mCherry</i>, or <i>emc6-GFP</i>, or co-transfected with <i>emc3-RFP/emc6-GFP</i>, <i>emc5-mCherry/emc6-GFP</i>, <i>emc6-GFP/RFP-rab7</i>. EMC6 puncta are indicated by arrows. Scale bar is 5μm. <b>b</b> Flowchart of Tandem Mass Tags Labeling coupled with LC-MS/MS for the comparative analysis of protein levels in retina of <i>emc2</i> (<i>GMR-Gal4/UAS-emc2A</i><sup><i>RNAi</i></sup>), <i>emc4</i> (<i>GMR-Gal4/UAS-emc4</i><sup><i>RNAi</i></sup>), <i>emc5</i> (<i>GMR-Gal4/UAS-emc5</i><sup><i>RNAi</i></sup>), and <i>emc7</i> (<i>FRT42D emc7</i><sup><i>1</i></sup>) mutant flies, as well as wild-type (<i>GMR-gal4</i>) flies. <b>c</b> Venn diagrams showing the overlap of proteins that were downregulated&gt;50% among the four analyzed genotypes: <i>emc2</i>, <i>emc4</i>, <i>emc5</i>, and <i>emc7</i></div>
</figcaption>
</figure>
<figure class="figure-item "
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itemprop="associatedMedia"
data-slide-index="4"
data-label-slug="fig-5">
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itemprop="contentUrl"
aria-describedby="figure-caption-4"
role="button"
data-image-width="700"
data-image-height="506"
data-image-alt="Fig. 5"
data-pmc-id="PMC7206144"
data-figure-id="Fig5">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-4"
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alt="Fig. 5" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="700">
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<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Fig. 5 </p>
</strong>
<div class="figure-caption-contents"><p> Retinal degeneration in <i> emc3 </i> ,… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 5 </p>
</strong>
<div class="figure-caption-contents"><p> Retinal degeneration in <i> emc3 </i> , <i> emc5 </i> , and <i> emc6 </i> mutant photoreceptors. TEM images… </p></div>
</div>
<figcaption id="figure-caption-4" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 5
</strong>
<div class="figure-caption-contents">Retinal degeneration in <i>emc3</i>, <i>emc5</i>, and <i>emc6</i> mutant photoreceptors. TEM images of representative ommatidia from (<b>a</b>) wild-type, (<b>b</b>) <i>emc3</i><sup><i>G7</i></sup>, (<b>c</b>) <i>emc5</i><sup><i>G2</i></sup>, and (<b>d</b>) <i>emc6</i><sup><i>N10</i></sup> flies. Flies were raised in a 12h-light/12h-dark (L/D) cycle for indicated time points. The degenerating photoreceptor cells are indicated by arrows. Scale bars are 2μm</div>
</figcaption>
</figure>
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itemprop="associatedMedia"
data-slide-index="5"
data-label-slug="fig-6">
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itemprop="contentUrl"
aria-describedby="figure-caption-5"
role="button"
data-image-width="750"
data-image-height="606"
data-image-alt="Fig. 6"
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data-figure-id="Fig6">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-5"
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</a>
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<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Fig. 6 </p>
</strong>
<div class="figure-caption-contents"><p> Loss of <i> Emc3 </i> in mammalian… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 6 </p>
</strong>
<div class="figure-caption-contents"><p> Loss of <i> Emc3 </i> in mammalian cone cells led to cone cell death. <b> a… </b> </p></div>
</div>
<figcaption id="figure-caption-5" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 6
</strong>
<div class="figure-caption-contents">Loss of <i>Emc3</i> in mammalian cone cells led to cone cell death. <b>a</b>, <b>b</b> Retina cryosections from (<b>a</b>) WT and (<b>b</b>) cone-KO littermate mice at P30. Sections were immunostained for M-Opsin (green) and the cone marker, peanut agglutinin (PNA, red). Cone cell number did not decrease at P30. <b>c</b>, <b>d</b> Retina cryosections from (<b>c</b>) WT and (<b>d</b>) cone-KO mice at P50. Sections were immunostained for M-Opsin (green) and PNA (red). There were almost no cone cells in cone-KO retinas. Nuclei were counterstained with DAPI. <i>ONL</i> outer nuclear layer, <i>OS</i> outer segment, <i>IS</i> inner segment. Scale bar is 25μm</div>
</figcaption>
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<figure class="figure-item tail"
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data-slide-index="6"
data-label-slug="fig-7">
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<strong class="figure-label">
<p> Fig. 7 </p>
</strong>
<div class="figure-caption-contents"><p> Retinal degeneration and RHO mislocalization… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 7 </p>
</strong>
<div class="figure-caption-contents"><p> Retinal degeneration and RHO mislocalization in <i> Emc3 </i> inducible KO mice. <b> a </b> Paraffin sections… </p></div>
</div>
<figcaption id="figure-caption-6" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 7
</strong>
<div class="figure-caption-contents">Retinal degeneration and RHO mislocalization in <i>Emc3</i> inducible KO mice. <b>a</b> Paraffin sections of mice retinas 11 days after induction (P31). Sections were stained with H&amp;E, and quantification of outer nuclear layer is shown on the left. Scale bar is 50μm. H&amp;E staining of paraffin sections and quantification of outer nuclear layer (ONL) nuclei revealed that the mutant ONL was reduced by ~50% compared to controls 11 days after induction. On top is an illustration of the whole mouse retina. <b>b</b> Immunofluorescence labeling of retina cryosections for control (WT) and mutant (KO) littermates at P31. Sections were labeled using antibodies against RHO (green) and DAPI (blue). NaK ATPase antibodies were used to label the inner segment. RHO was mislocalized to the inner segment and cell bodies in KO retinas (arrows). Scale bar is 25μm</div>
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