Fig. 6: NDUFA1 regulates cisplatin-induced renal tubular epithelial cell injury through modulation of FSP1.

A Immunoprecipitation confirmed the direct interaction between NDUFA1 and FSP1. HEK293T cells were co-transfected with Flag-FSP1 and HA-NDUFA1, followed by immunoprecipitation using anti-Flag M2 beads to pull down Flag-FSP1. The results demonstrated a direct interaction between NDUFA1 and FSP1. B Proximity ligation assay (PLA) detected the physical interaction between FSP1 and NDUFA1 in tubular epithelial cells after 24 h of cisplatin treatment. Red dots indicate positive PLA signals. Nuclei were counterstained with Hoechst. Scale bar = 10 μm. C Confocal laser scanning microscopy was used to observe the co-localization of FSP1 and NDUFA1 in tubular epithelial cells after 24 h of cisplatin treatment. Nuclei were stained with Hoechst for visualization. Scale bar = 10 μm. D PTCs were isolated from Idh1^WT/WTKsp^Cre and Idh1^WT/MutKsp^Cre mice. After PTCs infected with empty vector or Flag-Fsp1 lentivirus. 25 μm cisplatin were added. Representative PI staining images indicated cisplatin-induced PTC death. Scale bar = 100 μm. E Quantification of cell death using PI/Hoechst staining and Image J software. F PTCs were stained with Mitotracker and MitoSOX probes. Scale bar = 10 μm. G ROS generation and lipid ROS levels were detected using MitoSOX, DCFH-DA and C11-BODIPY probes in PTCs.