Phosphorylated p62 mediates AMPK-induced mitophagy in I(−) HCN cells.
A, co-transfection of Sh-rp62 HCN cells with plasmids encoding mt-mKeima and HA-tagged hp62-WT, -S294A, or -S294E. Cells expressing hp62-WT or hp62-S294E underwent mitophagy, whereas the expression of hp62-S294A mutant prevented mitophagy. Ex, excitation. B, analysis of A. The relative intensity ratio as an index of mitophagy (n = 8) is shown. C, a schematic diagram illustrating p62 phosphorylation by AMPK and subsequent mitophagy and ACD in HCN cells following insulin withdrawal. Error bars represent ±S.D. from independent assays. *, p < 0.05; ***, p < 0.001. Scale bar, 10 μm.