Fig. 7. Prevention of H₂O₂-induced apoptosis by I6CA was dependent on the activation of the Nrf2/HO-1 signaling pathway in V79-4 cells. The cells were treated with 300 μM I6CA or 10 μM ZnPP for 1 h and then treated with or without 1 mM H₂O₂ for an additional 1 h (A and B) or 24 h (C and D). (A) The medium was removed, and the cells were stained with DCF-DA. ROS production was measured using a flow cytometer, and representative profiles are shown. (C) The cells were stained with annexin V-FITC and PI for flow cytometry analysis, and representative profiles are presented. The percentages of apoptotic cells were determined by counting the percentages of annexin V-positive cells. (B and D) The data are expressed as the mean ± SD obtained from three independent experiments (***p<0.001 compared with the control group; ###p<0.001 compared with the H₂O₂-treated group; &&&p<0.001 compared with the I6CA and H₂O₂-treated group).