Fig. 4

ROS/ERK1/2 signals were involved in suppression effects of G-1 on CRC cell growth. (a) HCT-116 and SW480 cells were treated with various concentrations of G-1 for 3 h, and then loaded with CM-H2DCFDA. The fluorescence intensity was measured by FCM; (b) HCT-116 cells were pretreated with NAC (20 mM) for 1 h and then treated with 1 μM G-1 for 3 h, and then loaded with CM-H2DCFDA; (c) HCT-116 or SW480 cells were pretreated with NAC (20 mM) for 1 h and then treated with 1 μM G-1 for 48 h, cell viability was assessed by CCK-8 kit; (d) HCT-116 cells were treated with indicated concentrations of G-1 for 48 h with or without NAC pretreated for 1 h, and the protein expression was examined by western blot analysis; (e) HCT-116 cells were treated with 1 μM G-1 for 0 to 60 min, the total and phosphorylation of MAPK and Akt were measured by western blot analysis; (f) HCT-116 cells were pretreated with NAC (20 mM) for 1 h and then treated with 1 μM G-1 for 30 min, the protein expression was examined by western blot analysis; (g) HCT-116 cells were pretreated with 10 μM ERK1/2 inhibitor PD98059(PD), JNK inhibitor SP600125 (SP), or p38-MAPK inhibitor SB203580 (SB), and then exposed to 1 μM G-1 for 48 h, the cell viability was measured by use of CCK-8 kit. Data were presented as means ± SD of three independent experiments. **p < 0.01 compared with G-1 group