Extended Data Fig. 6: The acetylation-defective 6KR mutation of MED1 enhances chromatin occupancy of RNA Pol II which is also associated with endogenous acetylation level.
From: MED1 IDR deacetylation controls stress responsive genes through RNA Pol II recruitment
a, IGV snapshots of representative stress-induced genes with occupancy of HA (MED1) and Pol II (RPB1 S5P) shown for WT/KO and 6KR/KO cells. b, Comparable levels of representative Mediator subunits detected for association with immunoprecipitated endogenous MED17 in WT/KO, 6KR/KO and 6KQ/KO MCF7 cells. IgG was used as the IP control. c, Workflow depicting sequential ChIP-seq by anti-FLAG antibodies and pan-anti-acetyl-lysine antibodies to detect acetylation levels of FLAG-tagged MED1 (and its associated proteins) in genomic locations. d, Metaplots of occupancy of FLAG and acetylation for pre-defined MED1 peaks (from ChIP-seq in untreated MCF7) in High Ac, Mid Ac and Low Ac groups. e, Range-based heatmap of the occupancy of FLAG-MED1, acetyl-lysine (2nd ChIP), MED17, Pol II (RPB1) and p300 at pre-defined MED1 peaks (from ChIP-seq in untreated MCF7) that are stratified into 3 tiers based on relative acetyl-lysine density (lower 25%, middle 50% and upper 75% percentile, referred to as Low Ac, Mid Ac, High Ac). f, Metaplots of occupancy of MED17, Pol II (RPB1 S5P) and p300 for pre-defined MED1 peaks (from ChIP-seq in untreated MCF7) with comparison among High Ac, Mid Ac and Low Ac groups. g, h, Pie diagram showing the constitution of High/Mid/Low Ac groups of MED1 peaks for occupancy of MED17 and Pol II (g), and types of regulatory elements (h).
