Figure 7. SH003-induced p62 accumulation causes ROS-mediated apoptotic cell death.

(A) MDA-MB-231 cells were pretreated with or without NAC (2.5 mM) for 1 hour, followed by exposure to SH003. After 24 hours, cells were stained with H₂DCFDA for 1 hour at 37°C. ROS generation was detected with using FACSCalibur by the FL1 channel. (B) Cells were transfected with control siRNA and p62 siRNA and then treated with SH003 for 24 hours. ROS generation was measured by FACSCalibur. *P < 0.05. (C) Cells were pretreated with NAC for 1 hour and then treated with 500 μg/ml of SH003 for 48 hours. Cells were stained with annexin V and 7AAD. *P < 0.05. Experiments were performed in triplicate. Bars indicate means that standard deviations (SD).