Fig. 3. Huwe1 deficiency in hepatocytes aggravates ferroptosis in acute liver injury.

WT and Huwe1 HKO mice were subjected to I/R or CCl₄ injection. A Acsl4, Gpx4, Fsp1, DHODH and Cyp2e1 were detected by western blotting, n = 2-4; (B) Cyp2e1 protein levels were normalized to those of Gapdh, n = 3-4. C The mRNA level of Cyp2e1 was determined by real-time quantitative PCR in WT and Huwe1 HKO mice subjected to CCl₄ for 12 or 24 h. D Acsl4 protein levels were normalized to those of Gapdh, n = 3-4. E–F Representative images of liver sections stained with 4-HNE in WT and Huwe1 HKO mice subjected to I/R, (E) and CCl₄ for 12 or 24 h (F). Scale bars, 50 μm. G MDA levels were measured. n = 3–5. H–I mRNA levels of cytokines (TNFα, IL6 and IL1β) in WT and Huwe1 HKO liver lysates subjected to CCl₄ for 12 h (H) and I/R (I). n = 3–5. *p < 0.05, **p < 0.01, ***p < 0.001 compared in the indicated groups using unpaired two-tailed Student’s t-test.