Figure 5

Correlation between clusterin expression and AMPK activation. (A,B) Western blotting analysis of AMPK phosphorylation in the liver and eWAT of wild type and wCLU-tg mice fed CHOW or WD for 15 weeks. GAPDH was used as an internal control and was used to determine the relative ratio of AMPK phosphorylation. (C,D) Huh7 cells were incubated in 6-well plates. After 24 h, cells were incubated in serum-free DMEM. AMPK phosphorylation increased in a time- (0, 0.5, 1, 4, 12, 24 h) and dose- (0, 0.1, 0.5, 1, 2, 4 µg/ml) dependent manner in Huh7 cells. (E) Huh7 cells were pretreated with ML385 at 1, 2, 5, and 10 µM concentrations for 1 h and then treated with clusterin (2 µg/ml) for 48 h. (F) Huh7 cells were pretreated with Compound C at concentrations of 1, 2, 5, and 10 µM for 1 h and then treated with clusterin (2 µg/ml) for 48 h. (G) Huh7 cells were pretreated with TLR4 inhibitors, CLI-095 (1, 5 µM), and OxPAPC (15, 30 µg/ml), for 30 min and then treated with clusterin (2 µg/ml) for 24 h. *P < 0.05; **P < 0.01; ***P < 0.001.