Fig. 7

Irisin attenuates ferroptosis through the SIRT1/p53 Pathway. A SIRT1 protein expression in the heart was detected by western blotting (A1) and quantitative analysis (A2), All results were normalized to the expression level of GAPDH (n = 4 per group). B Representative Western blots in each of the four groups of H9C2 cells (B1) and quantification (B2) of the protein levels of SIRT1 in H9C2 cells, All results were normalized to the expression level of GAPDH (n = 4 per group). C Western blot images (C1) and quantification (C2) in each of the four groups of p53 K382 acetylation and p53 protein in H9C2 cells (n = 4 per group). D1 Representative fluorescent images of FerroOrange staining in each of the five groups of H9C2 cells (scale bar = 50 μm) and (D2) quantification of the FerroOrange fluorescence intensity (n = 6 per group). E1 Representative images of M-mode echocardiography of a mouse in each of the four groups of mice. Statistical analyses of (E2) LVEF%, (E3) LVFS%, (E4) LVIDd, and (E5) LVIDs in the indicated groups (n = 6 per group). F Representative TEM images of the heart tissue from a mouse in each of the four groups of mice (F1) and corresponding relative Flameng scores (Bottom) (F2) for mitochondria cardiac tissue (Scale bars = 500 nm; n = 5 per group). G Representative images of iron staining of the heart from a mouse in each of the four groups of mice (G1) and quantitative analysis (G2) of iron deposition in the four groups of mice (Scale bars = 20 μm; n = 5 per group). H p53 K382 acetylation levels and p53 protein levels were determined by Western blotting of the heart from a mouse in each of the four groups of mice (n = 4). Data are expressed as the mean ± SD. One-way ANOVA, and Bonferroni’s post-hoc test. ^*P < 0.05, ^**P < 0.01. HG high glucose, SIRT1 Sirtuin 1, STZ streptozotocin, LVEF left ventricular ejection fraction, LVFS left ventricular fractional shortening, LVIDd left ventricular internal diameter at end-diastole, LVIDs left ventricular internal diameter at end-systole, and TEM transmission electron microscopy