Supplementary Figure 5: ChIP assays to detect binding of BRD4, CDK9 and Pol II over the Myc gene loci.
From: BRD4 assists elongation of both coding and enhancer RNAs by interacting with acetylated histones
a, Effects of BRD4 knockdown by BRD4 shRNA on BRD4 protein and Myc transcript levels in NIH3T3 cells. b, Genome browser views of ChIP-seq signals for BRD4, CDK9 and Pol II (pan) along the Myc gene in NIH3T3 cells treated as indicated on the left. c, BRD4 knockdown cells were reconstituted with YFP-BRD4, YFP-BRD4-mBD or YFP alone. ChIP-qPCR assays using anti-YFP (GFP) antibody demonstrate binding of YFP-BRD4 or YFP-BRD4-mBD on the Myc gene locus. Signals relative to input DNA (y-axis) are plotted against positions of PCR primers (x-axis: covering the same region as the Myc gene locus shown at the bottom). In the gene locus diagram, dotted lines represent the upstream and downstream regions flanking the Myc gene. Background signals were assessed with cells expressing YFP alone. (d, e) ChIP-qPCR assays demonstrating binding of BRD4 (d) and CDK9 (e) on the Myc gene locus (shown at the bottom) in cells treated with JQ1 (1 μM, 2 h) or vehicle (DMSO). Background signals were assessed using normal rabbit IgG.
