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. 2017 Mar 31;8(18):30438–30454. doi: 10.18632/oncotarget.16737

Figure 7. The role of AMPK in A77 1726-induced autophagy.

Figure 7

(A & B) S6K1 knockdown and PF-4708671 induces AMPK and ULK1 phosphorylation. (A) A375 cells were transfected with S6K1 siRNA. After incubation for 48 hr, the cells were harvested and analyzed for AMPK and ULK1 phosphorylation with indicated antibodies. (B) A375 cells were treated with the indicated concentrations of PF-4708671 for 16 hr and analyzed for AMPK and ULK1 phosphorylation. (C) Inhibition of AMPK and ULK1 phosphorylation and LC3 expression by compound C (CC). A375 cells were incubated in the absence or presence of A77 1726 (200 μM) and/or CC (1 μM) for 16 hr. Cell lysates were prepared and analyzed for ULK and AMPK phosphorylation, and for LC3 and actin expression by Western blot. (D) The effect of AMPK activators on AMPK and ULK1 phosphorylation and LC3 expression. A375 cells were incubated in the absence or presence of A77 1726 (200 μM), oligomycin (10 μM) or metformin (5 mM) for 16 hr. Cell lysates were prepared and analyzed by Western blot with the indicated antibodies.