Fig. 2

DIG induced autophagy in liver cancer cells. (A and B) Transmission electron microscopy was used to detect autophagy in Hep3B and Huh7 cells that had been exposed to DIG at the recommended concentration for 48 h, scale bar = 5 μm. (C and D) Hep3B and Huh7 cells were pretreated with 3MA, AcDEVD, CHO, and Fer-1 inhibitors for 1 h, then treated with DIG for 48 h for CCK8 detection. (E-G) Expression of autophagy-related proteins in hepatoma cells treated with DIG, as detected by western blotting. (H and I) Immunofluorescence detection of LC3 autophagosome expression in cancer cells treated with DIG (×20). LC3B expression (green) was significantly increased. The nucleus is defined by DAPI staining (blue )( scale bar = 50 μm). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 compared with the control group. NS, not significant; Con, control; Fer-1, ferrostatin-1; 3MA, 3-methyladenine; DIG, digoxigenin