Figure 4

Triphala causes ROS generation in Capan-2 cells. A) Cells treated with 60 μg/ml Triphala for indicated time points were analyzed for DCF fluorescence (ROS generation) by flow cytometry after staining the cells with DCFDA. These experiments were repeated twice and obtained similar results. B) Activation of ERK, p53 and apoptosis by ROS can be abrogated by NAC. Cells were pretreated with 5 mM NAC for 1 hour and then treated with 60 μg/ml Triphala for 4 h. p-ERK, p-p53 and cleaved PARP were determined by western blotting as described in the method section. The blot was stripped and reprobed with anti-actin antibody to ensure equal protein loading. C) Effect of NAC on the transcriptional activity of p53. D) Apoptosis in NAC and Triphala treated cells was determined using cell death detection kit. Statistical significance in the treated group compared with control was analyzed by one-way ANOVA followed by Bonferroni's post hoc analysis. *Significantly different compared with PBS-treated control (P < 0.05). **Significantly different compared with Triphala treatment (P < 0.05).