Fig. 3

G-Rh2 regulates apoptosis through the mitochondrial-AIF pathway under serum-starved conditions. a Flow cytometric analysis of apoptotic HeLa and C-33A cells with DMSO or G-Rh2 under serum-free conditions in the presence or absence of 40 μM Ac-DEVD-CHO. b Flow cytometry in combination with Rhodamine 123 staining performed in HeLa and C-33A cells. Cells were treated with DMSO or G-Rh2 in the presence or absence of serum. c Subcellular distribution of cytochrome c and AIF in HeLa and C-33A cells. Cells were treated with DMSO or G-Rh2 in the presence or absence of serum for 24 h, and then collected for subcellular fractionation. COX IV, Tubulin, and Lamin B1 served as controls for mitochondria, cytoplasm, and nuclear fractions, respectively. Lane 1, 10% FBS and DMSO; lane 2, 10% FBS and 5 μM G-Rh2; lane 3, serum-free and DMSO; lane 4, serum-free and 5 μM G-Rh2. d Immunofluorescence analysis of AIF in cervical cancer cells. HeLa and C-33A cells were treated with DMSO or G-Rh2 in the presence or absence serum. DAPI was used for nuclear staining. G-Rh2 promotes apoptosis through an autophagy-dependent mechanism. Flow cytometric analyses of apoptotic HeLa cells with DMSO or G-Rh2 under serum-free conditions in the presence or absence of 100 nM BA1, 500 nM Rapamycin, or 5 mM 3-MA