In vivo antitumor activity of Triphala in a xenograft model. About 1 × 106 Capan-2 cells were injected subcutaneously on both left and right flanks of each athymic nude mouse and randomly assigned to three groups with 5 mice each group. Mice were fed with 50 mg/kg, or 100 mg/kg Triphala, 5 days/week by oral gavage. Control animals received PBS only. Tumors were measured by vernier calipers three times a week (Monday, Wednesday and Friday) once each mouse had palpable tumors. Each mouse was weighed twice a week (Monday and Friday). A) Average tumor volume in control (■), 50 mg/kg Triphala (▼) and 100 mg/kg Triphala treatment (◆). B) Average body weights of control and Triphala treated mice. Each data represents mean ± S.E. Error bars represent 95% confidence interval. *Statistically significantly different compared with control as analyzed by one-way ANOVA followed by Dunnett's test (P < 0.05). C) Tumor sections from control and Triphala treated mice were processed for H&E staining and immunohistochemistry for TUNEL, p-ERK and p-p53 staining. Each section was analyzed under the microscope at 200 × magnification and the number of stain positive cells were counted from three independent places on the slide. D) Immunoblot analysis for the expression of p-ERK (Thr-202/Tyr-204), p-p53 (Ser-15), ERK, and p53 using tumor lysates from control and Triphala treated mice. The blots were stripped and reprobed with actin antibody for equal protein loading. The numbers are the numerical representation of the mean densitometry values normalized to actin. These experiments were performed twice independently with similar results. Statistical significance was determined by one-way ANOVA followed by Bonferroni's post hoc analysis for comparisons.