Fig. 4

SIRT1 upregulation participated in ferulic acid-induced autophagy. a AML-12 cells were treated with different concentrations (0, 25, 50, and 100 μM) of ferulic acid (FA) for 12 h. Total cell lysates were probed by immunoblotting for SIRT1, phosphorylated AMPK, and phosphorylated Akt. b Cells were treated with 100 μM FA for 0, 4, and 8 h. SIRT1 protein abundance was detected. c Nuclear protein acetylation was detected after FA (100 μM) treatment for 12 h. d FA (100 μM) was added 2 h before 0.5 mM palmitic acid (PA) exposure. After 12 h of incubation, SIRT1 was detected by immunoblotting. e, f AML-12 cells were transfected with si-SIRT1 or scramble siRNA. The silencing efficiency was verified by SIRT1 mRNA and protein expression. g Autophagy-related proteins, including ATG5, ATG7, Beclin1 and LC3, were detected by immunoblotting. All values are shown as the means ± SD from three or more independent batches of cells. Bars with different superscripts are significantly different at p ˂ 0.05