Figure 4.

Erianin increases ROS generation and activates JNK signal pathway. (a) Cells were treated with increased concentrations of erianin 12 h, followed by loading with 10 μM of DCFH–DA for 30 min. The level of ROS was determined by fluorescence microscopy. (b) Cells were preincubated with NAC (5 mM) for 2 h and then treated with erianin for 12 h. Fluorescent intensity was detected using flow cytometry. Results are presented as mean±S.D. from three independent experiments. *P<0.05, significantly different compared with the control and NAC-treated group. (c) Cells were treated with various concentrations of erianin for 24 h. The expression of p-JNK, JNK, p-c-Jun, and Jun were analyzed by western blotting. (d and e) 143B was preincubated with SP600125 (30 μM) or NAC (5 mM) for 2 h and then treated with erianin for 24 h. Levels of p-JNK, JNK, p-c-Jun, and Jun were analyzed by western blotting