Fig. 6: The effect of sTim-3 and NF-κB inhibitors on cisplatin-induced mitochondrial oxidative stress in BUMPT cells.
From: Tim-3 protects against cisplatin nephrotoxicity by inhibiting NF-κB-mediated inflammation
After pre-treatment with PDTC or TPCA1 for 1 h, BUMPT cells were stimulated by 40 μM cisplatin with and without sTim-3 for 24 h. A Immunofluorescence analysis of calcium (green), 8-OHdG (green), and mitochondrial superoxide (MitoSOX) (red). The nuclei were labeled with DAPI (4′,6-diamidino-2-phenylindole, blue). Bar = 50 μm. B Mitochondrial membrane potential was visualized by the bright red fluorescence signals (JC-1 aggregates) and green signals (JC-1 monomer). Bar = 50 μm. C–F The fluorescence intensity of calcium, 8-OHdG, MitoSOX, and JC-1 were quantified using ImageJ software. G Measurement of total cellular reactive oxygen species (ROS) by flow cytometry; data from three independent experiments in BUMPT cells were processed by the GraphPad Prism software. H The mitochondrial membrane potential was visualized by JC-1 staining. Bar = 50 μm. I The fluorescence intensity of JC-1 was quantified by ImageJ software. Data represent mean ± SD (n = 4 per group). Statistical significance was determined using one-way ANOVA (G) or two-way ANOVA (C–F, I) followed by Tukey’s post hoc test. The experiments were repeated at least three times.
