Skip to main content
. 2021 Nov 19;49(21):12211–12233. doi: 10.1093/nar/gkab1122

Figure 2.

Figure 2.

CARM1 inhibitors eradicate BRD4 and me-BAF155 binding sites at SEs and decrease the expression of oncogenes addicted to SEs. (A) mRNA levels of me-BAF155 target genes (CDH1, CDCA7, COL1A2 and DDX18) after vehicle, JQ1, or TP-064 treatment in MDA-MB-468 cells measured by q-RT-PCR. **P< 0.01. (B) ChIP-qPCR analyses of me-BAF155 and BRD4 association with indicated genes after vehicle, JQ1, or TP-064 treatment in MDA-MB-468 cells. **P< 0.01; *P< 0.05; NS: not significant. (C) BRD4, me-BAF155 and H3K27Ac ChIP-seq signals in MDA-MB-468 cells after treatment with vehicle (black), JQ1 (red) or TP-064 (blue) in 2kb flanking regions of me-BAF155 peak summits from vehicle-treated cells. (D) ChIP-seq signals of me-BAF155, BRD4 and H3K27Ac in vehicle, JQ1 or TP-064 treatment conditions near HIF1A. (E) Super-enhancer signal-to-rank plots for MDA-MB-468 cells treated by vehicle, JQ1 or TP-064. The numbers of SEs were decreased from 184 to 15 and 31 by JQ1 and TP-064 treatment, respectively. (F) Western blotting of representative SE-regulated oncoproteins in vehicle, JQ1, TP-064, or JQ1 plus TP-064 treatment conditions in MDA-MB-468 cells. (G) BRD4 (top) and me-BAF155 (bottom) ChIP-seq signals before and after drug treatment. Signals were calculated in BRD4 (top) and me-BAF155 (bottom) peaks from MDA-MB-468 cells treated by vehicle. (H) Co-immunoprecipitation assays was performed using IgG, BRD4, me-BAF155, and BAF155 antibodies from nuclear lysates of MDA-MB-468 cells. Western blotting was performed using BRD4, me-BAF155, and BAF155 antibodies (top). The pull-down efficiency of each antibody was normalized by the band intensities of immunoprecipitates over the input using densitometry of the western blot bands (n = 3). (I) Decreased BRD4-association with BAF155 or me-BAF155 by increasing the amount of JQ1 in co-IP assays using MDA-MB-468 cells. Nuclear lysates were precipitated using BAF155 or me-BAF155 antibodies and co-precipitated BRD4 was detected by western blotting. (J) Proximity ligation assays showing the interaction between BRD4 and me-BAF155 was sensitive to JQ1 and TP-064 treatment in MDA-MB-468 cells.