Figure 5
Depletion of Survivin protects HeLa cells against docetaxel treatment. (a,b) HeLa cells were transfected with the indicated siRNAs, synchronized with single thymidine arrest, washed and released into fresh medium containing either DMSO or indicated concentrations of docetaxel for 24 h and analyzed for induction of apoptosis (a). Alternatively, synchronized cells were exposed to the indicated concentrations of docetaxel for 16 h, then stained with Hoechst 33,342 and photographed using phase and Hoechst fluorescence (b). (c,d) HeLa cells were synchronized with single thymidine arrest, washed and released into fresh medium containing the indicated concentrations of docetaxel in combination with either DMSO or Hesperadin (50 nM). Following 16 h combined treatment, cells were stained with Hoechst 33342 and photographed using phase and Hoechst fluorescence (c). Alternatively, following 24 h combined treatment, cells were collected and processed for assessment of apoptosis (d). (e) HeLa cells were transfected with the indicated siRNAs, synchronized with single thymidine arrest, released into medium containing the indicated concentrations of docetaxel for 24 h and analyzed for induction of apoptosis. (f) Cell viability assay. HeLa cells were incubated with the indicated siRNAs for 36 h followed by treatment with increasing concentrations of docetaxel for 48 h. Cell viability was determined by using the cck-8 reagent. The percentage of apoptotic or mitotic cells is shown. Values represent mean ± s.d. *Signifies p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant. Unpaired and two-tailed t test was used. Scale bars in (b) and (c), 50 μm.
