Supplementary Figure 8: Interchromatin granules in E2-regulated enhancer activity.
From: Phase separation of ligand-activated enhancers licenses cooperative chromosomal enhancer assembly
a. Effect of NRIP1 colocalization with ICG on its transcription b. Effect of TFF1 colocalization with ICG on its transcription. Data for a and b are median ± interquartile range. P-value was calculated using Wilcoxon rank sum test. >80 nuclei were examined in replicate c. Colocalization of transcribing TFF1 and NRIP1 loci with ICG. Highlighted area in square is magnified in image to the right. Arrowheads point to proximity of mTFF1 and mNRIP1 signals to the same ICG domain and arrows point to the weak NRIP1 signal that is not proximal to ICG. d. Transcriptional effect on NRIP1 locus colocalizing with a transcribing TFF1 locus in the same ICG. Data are mean ± s.d. P-value calculated by Mann-Whitney test. >100 nuclei were examined in replicate e. Knockdown efficiency of ICG component SRSF1and U2AF1 in MCF7 cells. siCTL represent scrambled oligo used for control transfection. Data pooled from 3 experiments. f. Q-PCR analysis demonstrated the inhibition of E2-induced eRNA expression from TFF1, NRIP1e, FOXC1e, and SMAD7e as a result of knockdown of SRSF1. g. Q-PCR analysis demonstrated the inhibition of E2-induced eRNA expression from TFF1, NRIP1e, FOXC1e, and SMAD7e as a result of knockdown of U2AF1. Data in e,f and g indicate mean ± s.d. P-value calculated using unpaired Student’s t-test. Pooled data from at least 3 independent experiments.
