Fig. 3: PTEC DNA damage causes mitochondrial dysfunction in cultured PTECs.

A Schematic diagram outlining the experimental design for cultured PTECs. Created in BioRender. https://BioRender.com/egn26ya (B) Real-time RT‒PCR analysis of biomarkers of renal proximal tubule injury and the rate-limiting enzyme involved in fatty acid β-oxidation in cultured PTECs overexpressing I-PpoI and controls. C OCR of cultured PTECs overexpressing I-PpoI and controls measured in a Seahorse XF24 system. Oligomycin, an inhibitor of ATP synthesis (1.5 μM), carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), a mitochondrial oxidative phosphorylation uncoupler (1.0 µM) and rotenone/antimycin A (0.5 μM), a complex I/III inhibitor, were injected sequentially at the indicated time points. D Basal and maximal OCRs, ATP production and storage respiration capacity in cultured PTECs overexpressing I-PpoI and controls. E OCR of cultured PTECs overexpressing I-PpoI and controls measured in a Seahorse XF24 system. Etomoxir, an irreversible inhibitor of CPT1 (4 µM), Oligomycin (1.5 μM), FCCP (1.0 µM) and rotenone/antimycin A (0.5 μM) were injected sequentially at the indicated time points. F Fatty acid oxidation (FAO) dependency in cultured PTECs overexpressing I-PpoI and controls. G Mitochondrial DNA copy number in the kidney and the liver. H Real-time RT‒PCR analysis of PGC-1α, a key regulator of mitochondrial biogenesis in the kidney and the liver. In A, G and H, n = 6 samples in each group. In B, E and F, n = 4 samples in each group. In C and D, n = 8 samples in each group. Each data point represents the mean ± SEMs. *p < 0.05, **p < 0.01 vs. the control. Source data are provided as a Source Data file.