Figure 6: dDsk2 regulates H2Bub1 deubiquitylation at promoters of dHP1c complex target genes.

(a) H2Bub1 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, ROW and WOC (red). Results are presented as fold enrichment with respect to the control (dsLacZ). Antibodies used were mouse monoclonal αH2Bub1 (N=5). (b) As in a, but for H2Bub1 levels on overexpression of full-length dDsk2 (light red) and the truncated ΔUBA–dDsk2 form (dark red). Results are presented as fold enrichment with respect to control cells expressing no construct (black; N=3). (c) As in a, but for H2Bub1 (top) and H3 (bottom) levels on depletion of dDsk2 (red), dUbp8/Nonstop (yellow) and both (blue; N=3). (d) mRNA levels of the indicated target genes are determined by RT–qPCR in S2 cells on depletion of dDsk2 (red), dUbp8/Nonstop (yellow) and both (blue). mRNA levels were determined in relation to control cells treated with dsRNA against LacZ (black). Results were normalized with respect to Actin5C levels (N=3). (e) As in a, but for ROW (top) and WOC (bottom) levels on depletion of dDsk2 (red), dUbp8/Nonstop (yellow) and both (blue; N=3). Error bars correspond to s.d.