Supplementary Figure 4: Effect of aliphatic alcohols on the E2 transcription program.
From: Phase separation of ligand-activated enhancers licenses cooperative chromosomal enhancer assembly
a. Q-PCR analysis of E2-induced eRNA expression from the TFF1, SMAD7 and PGR enhancer in MCF7 cells treated with indicated aliphatic alcohols (1,6-HD: 1,6-hexanediol, 2,5-HD: 2,5-hexanediol, 1,4-BD: 1,4-butanediol) compared to untreated cells (UT). Data indicate mean ± s.d. p-value calculated using unpaired Student’s t-test. Pooled data from at least 3 independent experiments b. Meta-analysis of GRO-seq data on MCF7 cells stimulated with E2 for 1 h followed by 5 min treatment with 2,5-HD or 1,6-HD on MegaTrans, Weak ERα and Non-ERα bound enhancer program genome-wide. c. Meta-analysis of GRO-seq data on MCF7 cells stimulated with E2 for 1 h followed by 5 min treatment with 2,5-HD or 1,6-HD on E2 regulated coding genes genome-wide. Boxes represent interquartile ranges (IQRs). Whisker represents points in lower and upper quartiles within 1.5 IQR from lower and upper edges of IQR d. Effect of 1,6-HD treatment on the recruitment of ERα, RARα and AP2γ on indicated enhancers. Results are shown as individual data points (circles), mean ± SD (lines). Data is representative of three independent experiments. e. Western blot on whole cell lystate from MCF7 cells treated with 7.5% of 2,5-HD or 1,6-HD for 5 min followed by 100 nM E2 for 30 min.
