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. 2022 Jul 6;13:930227. doi: 10.3389/fendo.2022.930227

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Copyright © 2022 Song, He, Indukuri, Huang, Stepanauskaite, Sinha, Haldosén, Zhao and Williams

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Generation and characterization of ERβ (only) MCF-7 cells. (A) Domain structure of ERα (upper) and genomic structure of ESR1 gene (bottom). The sgRNA target site is located at 5’-end of exon 1 and the 20-nt guide sequence is indicated in red letters. (B) ER expression was validated for the indicated receptors by Western blot analysis, using vinculin as loading control. Lane 1-2: ERβ (only) #8 and #32 clones, lane 3: ERα (only) cells (mock, in presence of Tet) and lane 4: showing induction of ERβ (mock, 20h after removal of Tet). (C) mRNA levels of IL20, pS2, GREB1 and PKIB were measured by qPCR in ERα (only) and ERβ (only, #32) following treatment with vehicle or E2. Data is represented as means + SD (n=3) and analyzed using two-way ANOVA followed by Bonferroni test, *P < 0.05, **P < 0.01, ***P < 0.001.