Figure 4.

Preventing ROS accumulation by antioxidant agent NAC reduced hirsutanol A-induced apoptosis. Cells were pre-incubated with NAC for 1h, then treated with hirsutanol A for 3h. The cellular H₂O₂ level was monitored by flow cytometry. Results are presented as means ± SD from 3 independent experiments. (** < 0.05 versus *; *** < 0.01 versus *) (A). Cells were pre-incubated with 1mmol/L NAC for 1h , then treated with various concentrations of hirsutanol A for 72h. The growth inhibition in SW620 and MDA-MB-231 cells were detected by MTT assay (B). Cells were pre-incubated with 1mmol/L NAC for 1h, then treated with various concentrations of hirsutanol A for 48h. Cell apoptosis was detected by AnnexinV/PI analysis (C).Cells were pre-incubated with 1mmol/L NAC for 1h, then treated with 20μmol/L hirsutanol A for 24h. The cleavage fraction of PARP was detected by immunoblotting assay (D).