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. 2021 Dec 8;12:7139. doi: 10.1038/s41467-021-27055-4

Fig. 5. BRD4 is necessary for e1 activity, but dispensable for the e1-SOX2 chromatin loop.

Fig. 5

a ChIP-seq profile of BRD4 and H3K27ac at the SOX2 locus in KYSE140 cells with 2 h of DMSO or 0.5 µM ARV-771 treatments. b The change of BRD4 ChIP-seq signal across all the BRD4 binding sites in KYSE140 cells after 2 h of 0.5 µM ARV-771 treatments, as compared to DMSO controls (two biological replicates). c RNA-seq results highlighting that SOX2 is one of the top genes downregulated in KYSE140 cells after 6 h of 0.5 µM ARV-771 treatments. d RT-qPCR results showing relative expression changes of SOX2 in six squamous cancer cell lines with 6 h of 0.1 or 0.5 µM ARV-771 treatments. The expression level was normalized to the DMSO controls. n = 2 biologically independent experiments. e RT-qPCR results showing relative expression changes of SOX2 in KYSE140 cells after two and 6 h of 0.5 µM ARV-771 treatments. The expression level was normalized to the DMSO control. n = 2 biologically independent experiments. f Left: schematic illustrating the 4sU labeling assay that was applied to capture nascent RNA. Right: RT-qPCR of the captured nascent RNA transcribed from the SOX2 gene in KYSE140 cells with 2 h treatments of DMSO or 0.5 µM ARV-771. RT-qPCR signal was normalized to the nascent RNA level of the HPRT1 gene and then to the DMSO control. n = 2 biologically independent experiments. g H3K27ac HiChIP loops that are connected to the SOX2 promoter in KYSE140 cells treated with 2 h of DMSO or 0.5 µM ARV-771. The color intensity corresponds to the number of PETs supporting each of the loops. h Schematic illustrating that ARV-771-mediated BRD4 degradation causes an acute and remarkable reduction in the e1 enhancer activity, but has little effect on the e1-SOX2 loop. Source data are provided as a Source Data file.