Supplementary Figure 10: Distinct effects of acute and chronic E2 treatment on transcription and biophysical properties of enhancer condensates.
From: Phase separation of ligand-activated enhancers licenses cooperative chromosomal enhancer assembly
a. ChIP experiment demonstrating the effect of acute and chronic E2 treatment on the recruitment of MegaTrans component RARα on TFF1 and NRIP1 enhancers. b. PRO-seq analysis on genes induced by 30 min of E2 treatment and transcriptional status of same set of genes after 14 h of E2 exposure c. PRO-seq analysis on genes transcribed by 14 h of E2 treatment and the transcriptional status of same set of genes after a second dose of 100 nM E2 for 1 h (after 14 h E2 treatment). For b and c, boxes represent interquartile ranges. Whiskers extend 1.5× the interquartile range from the 25th and 75th percentiles. P-values calculated with two sample t-test. d. Representative images of FRAP experiments quantified in Fig. 5b, showing ERα-mTurquoise foci formed after 30min (top panel) or 16 h (bottom panel) of E2 treatment, at indicated time points after bleaching. Photobleached regions are outlined in circles e. Representative image showing the effect of 1,6-HD on ERα/GATA3/TFF1 eRNA droplets formed in the presence of 5% PEG 8000 and 200 mM NaCl at indicated period after initial formation of the droplets.
