Fig. 7.

Fer-1 alleviated WBP2 knockdown-accelerated ferroptosis in CP-AKI. A Light microscopical analysis showed Fer-1 alleviated WBP2 knockdown-aggravated morphological shrinkage in cisplatin-treated BUMPT cells, DHE staining revealed that WBP2 knockdown-promoted ROS generation was attenuated by Fer-1; B H & E staining demonstrated that WBP2 knockdown-accentuated renal morphological disruption was partially restored by Fer-1 in cisplatin-treated kidneys, DHE staining revealed that Fer-1 mitigated WBP2 knockdown-accelerated ROS generation in cisplatin-treated kidneys; C - D CCK8 assay showed that RSL3 or cisplatin-induced cell death was accelerated by WBP2 knockdown in BUMPT cells, which was attenuated by Fer-1 treatment (n = 5; *P < 0.0001 compared with Lv-shNC group, ^#P < 0.0001 compared with Lv-shWBP2 group, 1-way ANOVA with Dunn's multiple comparisons); E Western blot studies demonstrated that WBP2-accelerated increase in NGAL and KIM-1 levels was mitigated by Fer-1 treatment in cisplatin-treated kidneys; F-G Western blot studies revealed that cisplatin-induced disruptions in ferroptosis-related markers (FTH1, GPX4 and NCOA4) was accentuated by WBP2 knockdown in BUMPT cells and kidneys, and these changes were attenuated by Fer-1 treatment. DMF: dimethylformamide, CP: cisplatin, Lv-shNC: empty vector lentivirus for WBP2 knockdown, Lv-shWBP2: lentivirus-mediated WBP2 knockdown; Ad-shNC: empty vector adenovirus for WBP2 knockdown, Ad-shWBP2: adenovirus-mediated WBP2 knockdown. Data are presented as mean ± SD. Scale bars: 100 μm.