Figure 3-1 Silibinin induced ROS ge

Figure 3-1 Silibinin induced ROS generation in HeLa cells via p53. The cells were cultured in medium with 10% fetal bovine serum, and then cultured in serum-free medium for 24 h. All following experiments in HeLa cells proceed according to this process. 1 he cells were treated with indicated inhibitors for 1 h, and cultured with 50 silibinin fbr 12 h. Con: control. Sili: silibinin. PFT-a and PFT: pifithrin-a. N : mole/liter. A: Inhibitory ratio v as detected in pifithrin-a (20 µM)-pretreated and silibinin-treated cells by MTT assay. B: Flow cytometric analysis was applied to determine the effect of PFT-a on silibinin-induced ROS generation in HeLa cells by staining with DCF-DA. C: Western blotting analysis was applied to detect the expression of p53/p-p53. a: The cells were treated with different concentration of silibinin (0, 25, 50 and 75 µM). b: The effect of PFT-a on p53/p-p53 expression was detected. D: The effect of NAC (2.5 mM) on the expression of p53/p-p53 was examined by Western blotting analysis.