Supplementary Figure 7: BRD4short facilitates progression of Pol II and transcript elongation on the Myc gene by binding to acetylated histones through bromodomains in vivo.
From: BRD4 assists elongation of both coding and enhancer RNAs by interacting with acetylated histones
a, Genome browser views of nascent transcript progression across the Myc gene locus, measured by chromatin RNA-seq analysis. b, BrU-labeled nuclear run-on RNA signals (measured by qRT-PCR) aligned along the Myc gene locus. Nuclei were isolated from BRD4 knockdown cells reconstituted with YFP-BRD4short, YFP-BRD4short-mBD or YFP alone, and subjected to a nuclear run-on reaction in the presence of 1% sarkosyl, and BrUTP, ATP, CTP, and GTP. The run-on RNAs labeled with BrU were fragmented, immunoprecipitated with anti-BrdU antibody, and analyzed by qRT-PCR using Myc locus specific primers. PCR efficiency was normalized with genomic DNA, and relative values of the run-on transcript signals are plotted against genomic positions corresponding to the PCR primers. (c, d) ChIP-qPCR assays for Ser2 phosphorylated Pol II (c) or K9/K14 acetylated H3 (d) along the Myc gene locus.
