Figure 2.

Senescence induced by loss of DPY30 correlates with activation of tumour suppressors of the INK4/ARF and CIP/KIP family. (A) mRNA expression levels of p21CIP1WAF1 (p21), p15INK4b (p15), and p16INK4a (p16) were monitored in control (shCtrl), H-Ras-expressing, or DPY30-depleted (shDPY30#1 and #3) IMR90 cells. Gene expression was quantified 6 days after selection by RT–PCR. Expression values were normalized to the housekeeping gene SF3B2 (SF3) and are illustrated relative to IMR90 control cells as fold expression (n=3). (B) p16INK4a (p16) and p15INK4b (p15) upregulation was measured by western blot using nuclear extracts of cells as in (A). Western blots were probed with a tubulin antibody to ensure equal loading. (C) Chromatin immunoprecipitation (ChIP) in the same cell types as in (A) 6 days after selection. DPY30, H3K4me3, and ETS1/2 were monitored at the ETS1/2 binding site within the promoter region of the p16INK4a gene. An unspecific IgG antibody was included as a negative control. H3K4me3 was normalized to histone H3. Values are presented as percentage of input (n=3).

Source data for this figure is available on the online supplementary information page.

Source Data for Figure 2 ^{(348.4KB, pdf)}