Fig. 6

ERβ and LRH-1 are part of the same transcriptional complex. A Proposed binding mechanisms on chromatin bound by both ERβ and LRH-1. B, C Illustration of intergenic and intronic binding sites of ERβ, with corresponding ChIP-seq peak (below), for sites also bound by LRH-1, by genes B Greb1, with distant to nearest TSS (isoform 1, NM_015764, or isoform 2, NM_001252071) indicated, along with the location of ERE and NR5A motifs, and C Cyp11a1, and Fkbp5. D ERβ ChIP-qPCR (n = 1, with 8 pooled ovaries from 4 mice) confirms binding of ERβ to respective site, normalized to input. E ERβ-LRH-1 ChIP-reChIP followed by qPCR (one out of two experiments illustrated here, with 14 pooled ovaries from 8 mice, in technical duplicates) supports dual binding of ERβ and LRH1-1 to Greb1 (intron 1), but not Cyp11a1 (intron 2) or Fkbp5 (intron 3) sites. Normalized to ERβ ChIP levels. F, G SW480 cells transfected to express ERβ and/or LRH-1, along with F ERE-TATA or G NR5A-RE luciferase reporter construct, show that ERβ and LRH-1 can repress each other’s transactivation activity (one out of three independent experiments per reporter construct illustrated here (n = 3), each with 3 technical replicates per condition, two-way ANOVA). # indicates significant difference by treatment (p < 0.05), ** p > 0.01, *** p < 0.001