Fig. 2: MDH2 regulates ccRCC proliferation through ferroptosis.

A Validation of MDH2 overexpression and knockout cell lines by Western blotting (NC: negative control for overexpression, OE: overexpression, KONC: negative control for knockout, KO1/KO2: knockout cell lines.). B MDH2 regulates the proliferation ability of normal renal epithelial cell lines. The statistical analysis comes from biological replicates and the experiment was repeated three times. Data was presented as mean ± SD. C MDH2 regulates the colony formation ability of normal renal epithelial cell lines. 50 visible colonies were observed on the 14th day for HK2 and HKC8. The statistical analysis comes from biological replicates and the experiment was repeated three times. Data was presented as mean ± SD. D MDH2 suppressed cell growth ability in ccRCC cells. The statistical analysis comes from biological replicates and the experiment was repeated three times. Data was presented as mean ± SD. E MDH2 suppressed colony formation ability in ccRCC. 50 visible colonies were observed on the 9th day for 786-O; on the 10th day for 769-P. The statistical analysis comes from biological replicates and the experiment was repeated three times. Data was presented as mean ± SD. F Treatment with regulated cell death inhibitors, including z-VAD-FMK (10 μmol/L), necrostatin-1 (20 μmol/L), bafilomycin A1 (100 nmol/L), VX-765 (5 μmol/L), liproxsrain-1 (10 μmol/L), and ferrostatin-1 (5 μmol/L), in 786-O MDH2-OE cells, and cell growth ability was evaluated by CCK-8. The statistical analysis comes from biological replicates and the experiment was repeated three times. Data was presented as mean ± SD. G ccRCC cells with 5 μmol/L ferrostatin-1, and cell growth ability and colony formation ability were evaluated. The statistical analysis comes from biological replicates and the experiment was repeated three times. Data was presented as mean ± SD. (*P < 0.05, **P < 0.01, ***P < 0.001, and ns not significant.).