Fig. 3: The IDH1-R132H mutation induces ferroptosis by promoting mitochondrial dysfunction.

A TECs (cell line) were infected with lentivirus encoding Flag-IDH1-WT and Flag-IDH1-R132H for 36 h, followed by stimulation with 25 μm cisplatin for 24 h. Western blot was performed to detect IDH1 expression after 36 h of infection. B, C TECs from (A) were seeded in 12-well plates. Colony counting was conducted on the 10th day after seeding. Cell proliferation was measured using the CCK-8 assay (C). D–I PTCs were isolated from Idh1^WT/WtKsp^Cre and Idh1^WT/MutKsp^Cre mice. D Representative PI staining images showing cisplatin-induced PTCs cell death. Scale bar = 100 μm. E Quantification of cell death using PI/Hoechst staining and Image J software. n = 6, ns not significant, ****P < 0.0001. F After 24 h of cisplatin treatment, cells were stained with Mitotracker and MitoSOX probes. Scale bar = 10 μm. G ROS production in Idh1^WT/WT and Idh1^WT/Mut PTCs induced by cisplatin was detected using DCFH-DA and C11-BODIPY probes, followed by analysis with flow cytometry. H Measurement of mitochondrial membrane potential (MMP) in IDH1^WT/WT and IDH1^WT/Mut cells after cisplatin treatment using JC-10 staining method. Results showed that the IDH1-R132H mutation exacerbated cisplatin-induced reduction in PTCs MMP. I Quantification of the ratio of red aggregates to green monomers. Scale bar = 10 μm.