Fig. 4.

Cd induces mitochondrial SIRT3 downregulation and GPX4 acetylation. CdCl₂ (2 mg/kg or 4 mg/kg) was injected intraperitoneally into adult BALB/c male mice. Kidney tissues were reserved 24 h after CdCl₂. (A–C) GPX4 and FSP1 were detected by immunoblot. (A) Representative images. (B) GPX4/β-actin; (C) FSP1/β-actin. (D–E) DHODH was detected by immunoblot. (D) Representative images. (E) DHODH/β-actin. (F–I) CdCl₂ (4 mg/kg) was injected intraperitoneally into adult BALB/c male mice. Kidney tissues were reserved 24 h after CdCl₂. Renal mitochondria and cytoplasm were extracted. (F and G) Cytoplasmic GPX4 was measured by immunoblot. (F) Representative images. (G) GPX4/β-actin. (H and I) Mitochondrial GPX4 was measured by immunoblot. (H) Representative images. (I) GPX4/VDAC1. (J and K) Mitochondrial SIRT3 was measured by immunoblot. (J) Representative images. (K) SIRT3/TOM20. (L–N) Mitochondrial GPX4 acetylation was analyzed by Co-IP. (L) Representative images. (M) Ace-Lysine binding efficiency. (N) GPX4/TOM20. (O) HK-2 cells were co-cultivated with CdCl₂ (10 μM) for 24 h. Co-localization of SIRT3, GPX4 and mitochondrial Tracker was observed by immunofluorescence. Red indicates SIRT3; Blue represents GPX4; Green represents Mito-Tracker. All data are presented as mean ± S.E.M. (N = 3–4). *P < 0.05, **P < 0.01. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)