{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T22:25:01Z","timestamp":1777415101952,"version":"3.51.4"},"reference-count":10,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2024,10,15]],"date-time":"2024-10-15T00:00:00Z","timestamp":1728950400000},"content-version":"vor","delay-in-days":14,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100006108","name":"National Center for Advancing Translational Sciences","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100006108","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2024,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Estrogen receptor alpha (ER\u03b1) is a nuclear receptor that is expressed mainly in the breast, uterus, and ovary, among several other organs. ER\u03b1 plays important roles in reproduction, mammary gland formation, and glucose homeostasis. Disruption of ER\u03b1 may result in adverse outcomes, such as cancer, impaired fertility, and abnormal fetal growth. Therefore, identifying compounds that modulate ER\u03b1 is of great interest due to their potential\u2010endocrine disrupting capability and pharmaceutical applications. To rapidly test tens of thousands of compounds, high\u2010throughput screening assays are essential. Here, we describe high\u2010throughput screening methods, including plating and treatment of cells in 384\u2010well and 1536\u2010well plates and analysis of the resulting data. The two cell lines used, MCF7\u2010VM7Luc4E2 and HEK293\u2010ER\u03b1\u2010bla, have been described previously. MCF7\u2010VM7Luc4E2 cells are a stable luciferase reporter gene cell line expressing full\u2010length endogenous estrogen receptor in the MCF7 cell line background, and HEK293\u2010ER\u03b1\u2010bla cells stably express an ER\u03b1 ligand\u2010binding domain\/GAL4 DNA\u2010binding domain fusion regulating a UAS \u03b2\u2010lactamase reporter gene. These cell lines can be used to identify and confirm ER\u03b1 modulators. Published 2024. This article is a U.S. Government work and is in the public domain in the USA. Current Protocols published by Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Establishment of a high\u2010throughput ER\u03b1 reporter gene assay with luminescence readout to identify activators and inhibitors of estrogen receptor \u03b1<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: Use of an orthogonal assay with fluorescence readout to confirm potential estrogen receptor activators or inhibitors<\/jats:p>","DOI":"10.1002\/cpz1.70029","type":"journal-article","created":{"date-parts":[[2024,10,15]],"date-time":"2024-10-15T10:49:10Z","timestamp":1728989350000},"source":"Crossref","is-referenced-by-count":1,"title":["Using Reporter Gene Assays to Screen and Identify Chemical Compounds that Modulate Estrogen Receptor Activity"],"prefix":"10.1002","volume":"4","author":[{"given":"Masato","family":"Ooka","sequence":"first","affiliation":[{"name":"National Center for Advancing Translational Sciences National Institutes of Health  Rockville Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Srilatha","family":"Sakamuru","sequence":"additional","affiliation":[{"name":"National Center for Advancing Translational Sciences National Institutes of Health  Rockville Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J. David","family":"Furlow","sequence":"additional","affiliation":[{"name":"University of California Davis  Davis California"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7285-8469","authenticated-orcid":false,"given":"Menghang","family":"Xia","sequence":"additional","affiliation":[{"name":"National Center for Advancing Translational Sciences National Institutes of Health  Rockville Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,10,15]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1177\/1087057115580298"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1097\/md.0000000000000211"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep05664"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijerph14030334"},{"key":"e_1_2_7_6_1","unstructured":"National Institute of Environmental Health Sciences (NIEHS). (2016).BG1Luc4E2 cells are being renamed VM7Luc4E2 cells. Retrieved fromhttps:\/\/ntp.niehs.nih.gov\/iccvam\/methods\/endocrine\/bg1luc\/bg1luc\u2010vm7luc\u2010june2016\u2010508.pdf"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.crtox.2022.100102"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.steroids.2014.06.012"},{"issue":"1","key":"e_1_2_7_9_1","first-page":"67","article-title":"Recombinant cell bioassays for endocrine disruptors: Development of a stably transfected human ovarian cell line for the detection of estrogenic and anti\u2010estrogenic chemicals","volume":"13","author":"Rogers J. M.","year":"2000","journal-title":"In Vitro & Molecular Toxicology"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a016709"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1093\/toxsci\/kfh180"}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70029","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,10,29]],"date-time":"2024-10-29T09:09:36Z","timestamp":1730192976000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.70029"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10]]},"references-count":10,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2024,10]]}},"alternative-id":["10.1002\/cpz1.70029"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.70029","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"value":"2691-1299","type":"print"},{"value":"2691-1299","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10]]},"article-number":"e70029"}}