Figure 3
From: Survivin safeguards chromosome numbers and protects from aneuploidy independently from p53
Increasing DNA contents after Survivin-RNAi are linked to a p53-dependent transient cell cycle arrest and to an attenuated entry into S-phase. a-c: Combined PI-staining of DNA and p21waf/cip in HCT116 and HCT116p53−/− cells 72 h after transduction of shRNA-vectors. a: Graph showing the mean values and SEM of p21waf/cip-positive cell fractions after knockdown of Survivin at different DNA contents ranging from 2n to greater than 8n (**p < 0.01). b: Representative FACS analysis showing p21waf/cip staining in HCT116 (wt) and HCT116p53−/− cells. c: Quantitative analysis of the induction of p21waf/cip in total HCT116 and HCT116p53−/− cell populations after transduction of shSurv and shLuc control, respectively (**p < 0.01; n = 4) d-f: Combined PI-staining of DNA and Cyclin D1 in HCT116 and HCT116p53−/− cells after transduction of shRNA-vectors. d: Graph showing the mean values and SEM of Cyclin D1-positive cell fractions after knockdown of Survivin at different DNA contents (**p < 0.01). e: Representative FACS dot blots showing Cyclin D1 analyses in HCT116 (wt) and HCT116p53−/− (p53−/−) cells f: Quantitative analysis of the induction of Cyclin D1 expression in total HCT116 and HCT116p53−/− cell populations after transduction of shSurv and shLuc control, respectively (**p < 0.01). g-i: BrdU-incorporation analysis. g: Graph showing the mean values and SEM of BrdU-positive cell fractions after knockdown of Survivin in S-phases of cells (S1 to S3). Note, that despite p21waf/cip and Cyclin D1 accumulations HCT116 cells with DNA content >4n are still able to incorporate BrdU. h: Representative FACS dot blots showing BrdU stainings after Survivin knockdown. i: Effect of Survivin knockdown on BrdU incorporation in total HCT116 and HCT116p53−/−cell populations and the corresponding shLuc controls. Data represents mean values and SEM (*p < 0.05; **p < 0.01).