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. 2020 Jun 5;6(23):eaaz7249. doi: 10.1126/sciadv.aaz7249

Fig. 2. Enhancer dynamics mediated by RING1B.

Fig. 2

(A) Western blots of histone modifications in control and RING1B-depleted cells in HD condition and upon E2 administration. Histones were extracted using sulfuric acid. (B) H3K27ac ChIP-seq signal across the right arm of the chromosome 17 in control and RING1B-depleted cells. (C) SEs identified in each E2 time point. (D) Venn diagram of SEs and genes associated with SEs in each E2 time point. (E) Genome browser screenshots of H3K27ac ChIP-seq in SEs identified in the HD and 24 hours of E2 condition (BCAM SE), only after 24 hours of E2 (GREB1 SE), and at all the time points analyzed (DSCAM SE). (F) H3K27ac signal in control and RING1B KD cells at sites that acquired H3K27ac after 24 hours of E2 in control cells. Significance was determined by Mann-Whitney U test. (G) Genome browser screenshots of H3K27ac in control and RING1B KD cells before and after 8 and 24 hours of E2. (H) RT-qPCR analyses of enhancer RNA expression at the GREB1 and E2F6 SEs in control and RING1B-depleted cells in the HD condition and after 24 hours of E2. mRNA expression was normalized to the housekeeping gene RPO. N = 3.