Figure 7: dDsk2 regulates RNA pol II pausing at promoters of dHP1c complex target genes.
From: dDsk2 regulates H2Bub1 and RNA polymerase II pausing at dHP1c complex target genes
(a) The levels of RPB3 (top) and the promoter-proximal Pol IIoser5 form (bottom) across selected target genes are determined by ChIP–qPCR at the indicated positions with respect to TSS in control S2 cells treated with dsRNA against LacZ (black) and on depletion of dDsk2 (red), dUbp8/Nonstop (yellow) and both (blue). Results are presented as fold enrichment with respect to the control (dsLacZ) at the most proximal position to the TSS. The position of the TSS is indicated. Antibodies used were rabbit polyclonal αRPB3 and αIIoser5 (N=3). (b) The relative abundance of RPB3 (black) and Pol IIoser5 (red) at TSS with respect to their total abundance across the gene is determined for the indicated target genes in control S2 cells treated with dsRNA against LacZ and on depletion of dDsk2, dUbp8/Nonstop and both (N=3). (c) NELF-E levels at the indicated positions with respect to TSS of selected target genes are determined by ChIP–qPCR in control S2 cells treated with dsRNA against LacZ (black) and on depletion of dDsk2 (red). Results are presented as fold enrichment with respect to the control (dsLacZ). Antibodies used were rabbit αNELF-E antibodies (N=3). (d) H3K4me3 (left), acetylated H3 and H4 (PanAc; centre) and acetylated H4 (AcH4; right) levels at the indicated positions with respect to TSS of selected target genes are determined by ChIP–qPCR in control S2 cells treated with dsRNA against LacZ (black) and on depletion of dDsk2 (red). Results are presented as fold enrichment with respect to the control (dsLacZ). Antibodies used were rabbit polyclonal αH3K4me3, αPanAc and αAcH4 (N=3). Error bars correspond to s.d.
