Figure 3.

ABC294640 increased TRAIL induced apoptosis. A549 and H1299 cells were grown on 6-well plates to 70-80% confluence and treated with indicated concentrations of TRAIL (100 ng /ml) or ABC294640 (75 mM) or both of the above treatments, or non-treatment. After 24 h exposure, (A) morphologic assessment were captured using Olympus IX70 Inverted Microscope at a magnification of 100, (B-E) cells were stained with PtdIns and Annexin V-FITC, then apoptosis were analyzed by flow cytometry as described in Materials and Methods. Data are presented as total apoptosis rate. Mean §SEM of 3 independent experiments. (F and G) caspase-3/8/9 and Bid were assessed by Western blot of cell lysates, as described in Materials and Methods. GAPDH expression served as a loading control. Representative of 3 experiments.