Fig 1: High SLC25A11 levels increased the mitochondrial membrane potential in cancer cells. (a) The mitochondrial membrane potential was analyzed by live cell imaging in cancer cell lines in comparison with normal cells using ZEN software (scale bar, 20 µm, 10 µm). (b) The oxygen consumption rate (OCR) was analyzed using the Seahorse XFe analyzer in cancer cell lines compared to normal cells and then normalized by SRB assay. (c) The expression levels of oxidative phosphorylation (OxPhos) components were analyzed in NSCLC and melanoma cells by immunoblotting. (d) Cytosolic/mitochondrial NADH ratio in cancer cell lines compared with normal cells. Western blot confirming the isolation of mitochondria from cytosol with MDH1 (cytosol marker) and CV-ATP5A (mitochondria marker) antibodies. (e) Malate-aspartate shuttle (MAS) for NADH transportation into the mitochondrial matrix. MAT, malate-a-ketoglutarate transporter; GAT, glutamate-aspartate transporter; OAA, oxaloacetate; a-KG, a-ketoglutarate. (f) The expression levels of SLC25A11, GOT1, GOT2, MDH1, and MDH2 in NSCLC and melanoma cells were analyzed by immunoblotting and quantified by ImageJ. (g) Comparison of SLC25A11 expression in cancer cells and normal cells by immunofluorescence staining and intensity analyzed by ZEN software (scale bar = 20 µm). (Data were presented as mean ± SD. ***p < .001, **p < .01, *p < .05).