Figure 3

LY294002 modulated oxaliplatin-induced cell apoptosis. (A) Cells were added with 10 μM LY294002, 0.5 μM RAD001 or control vehicle and then treated with 0–200 μM oxaliplatin. TUNEL assay was done as described in the Methods section. The percentage of apoptotic cells in each group of treatments was demonstrated (values shown as mean ± SD, *; p < 0.05 versus the same concentration of oxaliplatin). (B) Apoptotic cells (green spots) were detected with an ApopTag Staining kit and counterstained with DAPI (blue). RMCCA1 cells were treated with Vehicle (a) and 100 μM oxaliplatin (b). Cells were treated with RAD001 plus 100 μM oxaliplatin (c) and with LY294002 plus 100 μM oxaliplatin (d) (Arrow head indicates the apoptotic cells). (C) Western blots analysis of cleaved caspase-3 and caspase-3 in each group of treatments. The blots were representative of three independent experiments.