Fig. 3

2-APQC alleviates myocardial fibrosis to improve HF by SIRT3 activation in vivo. a, b Immunohistochemical detection of ac-MnSOD2 (K68) and ac-MnSOD2 (K122) in rat heart tissues in different groups. Data are present as mean ± s.e.m, n = 3. c The expression levels of Ac-MnSOD2 (K68), ac-MnSOD2 (K122), SIRT3 and acetylated lysine were detected by Western blot. d, f Immunohistochemical detection of α-SMA cells with α-SMA in rat heart tissues in different groups. The position of the nucleus is marked by DAPI staining. Scale bar = 25 μm. Data are present as mean±s.e.m, n = 3. e, g Immunohistochemical detection of masson staining in rat heart tissues in different groups. Data are present as mean±s.e.m, n = 3. h Detection of Akt/ mTOR/p70S6K pathway by Western blot. i Detection and quantification of the expression of TGF-β/Smad3 pathway, TGF-β, JNK, p-JNK, Smad3, p-Smad3 and lysyl oxidase (Lox) by Western blot. j The expression levels of Akt, p-Akt, JNK, p-JNK, α-SMA, fibronectin and collagen I in rat heart tissues in different groups were detected by Western blot. k H9c2 cells were treated with indicated concentration of 2-APQC in the presence of absence of nicotinamide for 24 h, and then treated with ISO for 48 h, the cell viability was measured by the MTT assay. l H9c2 cells were treated with indicated concentration of 2-APQC in the presence of absence of nicotinamide for 24 h, and then treated with ISO for 48 h. Determine the size of cardiomyocytes by β-actin tracker staining (red). The position of the nucleus is marked by DAPI staining. Scale bar = 10 μm. Data are present as mean ± s.e.m, n = 3. m SIRT3 knockdown plasmid transfection was conducted 48 h prior to cell treatment. H9c2 cells were treated with indicated concentration of 2-APQC for 24 h, and then treated with ISO for 48 h. Determine the size of cardiomyocytes by β-actin tracker staining (green). The position of the nucleus is marked by DAPI staining. Scale bar = 10 μm. Data are present as mean ± s.e.m, n = 3. n Detect the expression of p-AKT, p-mTOR, p-JNK and p-SMAD3 by Western blot. o Detect the expression of p-AKT, p-mTOR, p-JNK and p-SMAD3 by Western blot in SIRT3 KO mice. p Detect the expression of p-AKT, p-mTOR, TGF-β and p-SMAD3 by Western blot. q Detect the expression of JNK, p-JNK, α-SMA, fibronectin and collagen I by Western blot. ns, no significance, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, compared with ISO group; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001, ^####p < 0.0001, compared with control group. Nico nicotinamide, ISO isoproterenol