Figure 3. BBD24 induced multiple cell death pathways of HOS cells. Human osteosarcoma HOS cells were treated with BBD24 at 2 μg/ml for indicated times. The cells were harvested by trypsinization and collected by centrifugation for analysis of cell viability, apoptosis and necrosis using flow cytometry assay. HOS osteosarcoma cells were treated with BBD24 at the indicated concentrations for 48 hours, followed by Western blot analysis for caspase family and LC3-II. (A–C) Flow cytometry assay indicated that BBD24 induced apoptosis and necrosis of HOS cells with time dependence. (D–I) BBD24 promoted activation of caspases and autophagy in HOS cells in a dose-dependent manner. Results were expressed as means ± SD of three independent experiments. * P<0.05.