Extended Data Fig. 7: RB-3 regulates expression of target genes and impairs enrichment of H2Aub and RING1B on target gene promoter regions in TEX cells.
From: Small-molecule inhibitors targeting Polycomb repressive complex 1 RING domain
a, qRT–PCR showing time dependent changes in transcript levels of CD34, C/EBPα, CD-86 upon treatment with 25 μM RB-3 and RB-nc. Representative data out of two replicates. b, Analysis of the H2Aub and H3 levels, and binding of RING1B to the two promoter regions of C/EBPα in TEX cells treated with RB-nc and DMSO for 8 days using ChIP assay. Representative data out of two replicates. c, Analysis of the H2Aub and H3 levels, and binding of RING1B to the two promoter regions of CD34 in TEX cells treated with RB-3 (red), RB-nc (blue) and DMSO (black) for 8 days using ChIP assay. Representative data out of two replicates. d, Analysis of the H2Aub and H3 levels, and binding of RING1B to the two promoter regions of ITGAM in TEX cells treated with RB-3 (red), RB-nc (blue) and DMSO (black) for 8 days using ChIP assay. Representative data out of two replicates. Two promoter region primers between 1 to 2 kb upstream of the transcription start site (TSS) for each of CD34, C/EBPα and ITGAM promoter were selected and analyzed (panels b, c, d). e, Average genome-wide occupancy of H2Aub around the transcription start sites (TSS) and genomic regions in TEX cells determined from ChIP-seq experiment. TEX cells were treated with DMSO and 25 μM RB-3 for 6 days.
