Fig. 3

CHTM1 regulates AIF1 cellular distribution in response to metabolic stress. (a, left panel) Representative fluorescent photomicrographs showing increase in nucleo-cytosolic distribution of endogenous AIF1 (green) in CHTM1 knockdown A549 cells following glucose/glutamine starvation for 4 h. White arrows indicate cells with nuclear localization of AIF1. Scale bar, 10 μM. (Right panel) Quantitative results showing the relative numbers of A549 cells with cytosolic/nuclear/mitochondrial localization of AIF1 (n indicates the number of cells analyzed). (b) Representative Western blot showing increase in cytosolic AIF1 levels in CHTM1 knockdown MCF-7 breast cancer cells following 12-h glucose/glutamine starvation. (c) Representative Western blot showing decrease in levels of cytosolic AIF1 in CHTM1 overexpressing A549 lung cancer cells following 4-h glucose/glutamine starvation (compare lane 7 with 8). (d, left panel) Representative fluorescent photomicrographs showing increase in nucleo-cytosolic distribution of endogenous AIF1 (green) in CHTM1 knockdown A549 cells following 12-h metformin treatment. White arrows indicate cells with nuclear localization of AIF1. Scale bar, 10 μM. (Right panel) Quantitative results showing the relative numbers of A549 cells with cytosolic or nuclear localization of AIF1 following metformin treatment (n indicates the number of cells analyzed). (e) Representative Western blot analyses showing increase in AIF1 cytosolic levels in CHTM1 knockdown A549 cells upon 12-h metformin treatment (compare lane 10 with lanes 11&12)