Figure 6.

ROS generation is required for the activation of AFB1-induced autophagy. (A) RAW264.7 cells were pre-treated with the NOX2 inhibitor DPI (50 μM) for 1 h. PMA (16 nM) and AFB1 (0.25 μM) were then added to the cells for 2 h and western blotting of LC3 was performed. (B,C) RAW264.7 cells were treated with AFB1 at different concentrations and times. Cytosolic ROS were labeled by DHR 123 (1 μM) and were detected by a plate reader, ^**P < 0.01, ^***P < 0.001 compared with the control groups in the same cell line. (D) Cells were pretreated with wortmannin (100 nM, 1 h) after treatment with AFB1 (0.25 μM) and PMA (16 nM) for 2 h. Cytosolic ROS were detected by a plate reader, ^***P < 0.001 compared with the control groups in the same cell line.