Fig. 6

GRHL2 differential ChIP-seq between 0 and 45 min. (a) Activation of the ER with estro-2-diol results in a genome-wide increase in GRHL2 binding. (b) VULCAN analysis of the same data shows a significant enrichment for ESR1 sites in both the context of the METABRIC and TGCA networks. The regulon for FOXA1 is also not enriched. Inspection of known FOXA1/GRHL2 sites (e.g., RARa promoter) shows GRHL2 already bound. (c) Overlap of GRHL2 binding with public datasets shows that E2-responsive GRHL2 sites show considerable overlap with ER, FOXA1, and P300 sites; H3K4Me1 and H3K4Me3 show little enrichment. (d) Analysis of P300 binding showed a greater overlap of GRHL2 ER-responsive sites in the presence of E2 than in control conditions. (e) Overlap with ER ChIA-PET sites showed enrichment for GRHL2 sites at ER enhancers. (f) Analysis of Gro-SEQ data (GSE43836) at GRHL2 sites. Blue lines are control samples, pink lines are samples after stimulation with E2. In general, GRHL2 sites (left) show no change in the levels of transcription on the addition of E2; however, E2-responsive GRHL2 sites (right) show a robust increase in transcription on the activation of the ER. (g) Motif analysis of differentially bound sites gave the top two results as GRHL2 and ER