Effects of NVP-BEZ235 on survival of HMC-1 cells and KU812 cells.

<p>A,B: HMC-1.1 cells (left panels), HMC-1.2 cells (middle panels), and KU812 cells (right panels) were cultured in control medium (Co), LY294002 (20 µM), or increasing concentrations (0.001 µM–10 µM) of NVP-BEZ235 (A) or RAD001 (B) at 37°C for 48 hours. Then, the numbers (percentage) of apoptotic cells was counted on Giemsa-stained cytospin slides. Results represent the mean±S.D. of three independent experiments. Asterisk (*) indicates p<0.05. C: HMC-1.1 cells (upper panels), HMC-1.2 cells (middle panels) and KU812 cells (lower panels) were incubated in control medium (Co) (left panels) or in medium containing NVP-BEZ235, 1 µM (middle panels) or RAD001, 1 µM (right panel) for 48 hours. After incubation, cells were examined for combined AnnexinV/PI staining by flow cytometry. D: HMC-1.1 (left panel), HMC-1.2 (middle panel), and KU812 cells (right panel) were incubated in control medium (Co) or in various concentrations of NVP-BEZ235 or RAD001 as indicated (37°C, 48 hours). Then, the percentage of active caspase 3-positive cells was determined by flow cytometry. Results represent the mean±S.D. of three independent experiments. Asterisk (*) indicates p<0.05. E: HMC-1.1 cells (upper panels), HMC-1.2 cells (middle panels) and KU812 cells (lower panels) were incubated in control medium (Co) or in medium containing NVP-BEZ235 (1 or 10 µM) or RAD001 (1 or 10 µM) (as indicated) at 37°C for 48 hours. Thereafter, cells were harvested and subjected to Tunel assay and examined by fluorescence microscopy.