Fig. 5

SLC19A2 promoted the mitochondrial respiration of NONO-TFE3 rRCC by increasing the NAD⁺ transportation into mitochondria. A UOK109 cells were transfected with lentivirus shRNA (NC), shRNA (NMRK2), or shRNA (SLC19A2). The mitochondrial respiration of cells in each group was measured by the Seahorse XF Extracellular Flux Analyzer with a Seahorse XF Cell Mito Stress Kit. B, C And the NAD⁺ level of the whole cell or D, E of mitochondria of cells in each group was measured with an NAD⁺/NADH Assay Kit. F The subcellular localization of the SLC19A2 protein in UOK109 cells was observed by IF. G The mitochondria in UOK109, UOK120, and 786-O cells were isolated, and the total protein in the cytoplasm and mitochondria was extracted. The expression of SLC19A2 protein in cytoplasm and mitochondria was detected by Western Blotting. GAPDH and Tom20 were used as the positive control for periplasmic protein and mitochondrial protein, respectively. H, I UOK109 cells were transfected with lentivirus shRNA (NC), shRNA (NMRK2), or shRNA (SLC19A2). Then the cells in each group were treated or untreated with 100 μM NMN for 24 h. The NAD⁺ level in mitochondria was measured with an NAD.⁺/NADH Assay Kit. J, K And the mitochondrial respiration of cells in each group was measured by the Seahorse XF Extracellular Flux Analyzer with a Seahorse XF Cell Mito Stress Kit. Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001