Figure 3.

Metformin activated the autophagy signaling pathway in D407 cells. (a) Cells were transfected with tandem fluorescent-tagged LC3 reporter and treated with 60 μM CQ and 1 mM metformin plus 60 μM CQ for 2 h. Images were taken by a confocal microscope (scale bars, 20 μm). (b) D407 cells were treated with 60 μM CQ and 1 mM metformin plus 60 μM CQ for 2 h. The expression of LC3B-I and LC3B-II was assessed by Western blot. (c) D407 cells were treated with different concentrations of metformin for 2 h, and the expression of LC3B, p62, P-Beclin1, and P-ULK1 was assessed by Western blot. (d) D407 cells were treated with 1 mM metformin for the periods of time indicated. The expression of LC3B, p62, P-Beclin1, and P-ULK1 was assessed by Western blot.