Fig. 4

Knockout of Plin5 diminished HFHC diet-induced ferroptosis. A, B Total and ferrous iron levels in liver were measured by iron probe in WT and Plin5 KO mice fed an HFHC-diet. C Prussian blue iron staining (Enhanced with DAB) of liver sections from mice fed an HFHC-diet. Scale bar, 50 um. D, E Serum total and ferrous iron levels were measured by iron probe in WT and Plin5 KO mice fed an HFHC-diet. F Expression of hepatic iron metabolic genes, Tfr1, Hamp1, Hamp2, Fth, Ftl, was measured by qPCR from WT and Plin5 KO mice fed an HFHC-diet. G, H MDA and SOD content was measured using a malondialdehyde assay kit (TBA method) and total superoxide dismutase assay kit (hydroxylamine method) respectively. I Hepatic mRNA level of Hmox1, Ptgs2 and Nox2 was measured by qPCR in HFHC-diet fed mice. J Confocal images of WT and Plin5 KO mouse liver sections labeled with C11-BODIPY and DAPI from mice fed on HFHC-diet. Green and blue colors indicate lipid ROS (peroxidated lipids) and nucleus respectively. Scale bar, 50 um. K Western blot analysis of GPX4 in WT and Plin5 KO mouse livers. A two-tailed Student t-test was used for statistical analysis. The data are presented as the mean ± SEM. *< 0.05, **< 0.01. Abbreviations: MDA: malondialdehyde; SOD: total superoxide dismutase. (n = 6/group)