Fig. 6

Amifostine-induced AMPK activation is mediated by SIRT1. a Western blot detection of SIRT1 expression in human fibroblast cells stimulated by TGF-β1 or amifostine. n = 3. The results shown are cropped from the original blots. b SIRT1 activity assay was used to determine SIRT1 activity in human fibroblast cells after stimulation by TGF-β1 or amifostine treatment. SIRT1 activity is plotted as a fold-change compared to PBS control cells. n = 5, ***p < 0.001. c SIRT1 was inhibited by its specific inhibitor Ex-527 (5 µM, 30 min). Cells were then treated with amifostine. Activation of AMPK was detected with the Thr172 phosphorylated AMPK antibody. The results shown are cropped from the original blots. d Endogenous SIRT1 was downregulated by transfection with SIRT1 siRNA. Phosphorylation of AMPK at Thr172 was detected by Western blot. n = 3, *p < 0.05, ***p < 0.001, **** < 0.0001. The results shown are cropped from original blots. e Cells were treated with SIRT1 activator resveratrol (10 µM, 30 min) followed by TGF-β1 treatment. Phosphorylation of AMPK was determined by Western blot. n = 3, *p < 0.05, **p < 0.01, ***p < 0.001, **** < 0.0001