Figure 3
Os-induced cell death is ROS-dependent. (a) A total of 3 × 105 HCT116 cells were incubated with 50 μM of Os for 1 or 3 h, and intramitochondrial O2− was determined using the fluorescent dye MitoSox Red and analyzed by flow cytometry. (b) Cells were preincubated with 1.5 mM of NAC for 1.5 h before incubation with 50 μM of Os for 4 h, and intracellular H2O2 was detected by DCFH-DA loading and analyzed by flow cytometry. (c) Cells were preincubated with 1.5 mM of NAC for 1.5 h before incubation with 50 μM of Os for 24 h. Cell viability was determined by crystal violet. (d) Cells were treated with NAC and Os as in (b) and 20 000 cells were seeded onto 100-mm petridishes for assessment of colony formation. (e) After preincubation with NAC and Os treatment, cell lysates were used for western blot analysis using PARP antibody. ••P<0.025 versus control
