fig4

Figure 4. FASN inhibition attenuates TNNI3 p.R186Q-induced aberrant fatty acid metabolism and cardiac hypertrophy. Tnni3^R186Q/R186Q mice at 10 months of age were administered saline or C75 (15 mg/kg/day) daily for 8 weeks. (A) Experiment timeline for Tnni3^R186Q/R186Q mice. (B) Representative M-mode echocardiographic images of Tnni3^+/+, Tnni3^R186Q/R186Q and Tnni3^R186Q/R186Q +C75 mice. (C) Lipogenesis in the ventricular myocardium of Tnni3^+/+, Tnni3^R186Q/R186Q and Tnni3^R186Q/R186Q +C75 mice was determined by Oil Red O staining using frozen sections (n = 3 per group). (D) The levels of TG and TC were individually measured using enzyme-linked immunosorbent assay in the ventricular myocardium of Tnni3^+/+, Tnni3^R186Q/R186Q and Tnni3^R186Q/R186Q +C75 mice (n = 3 per group). (E) HE (pink) and Masson’s trichrome staining (red) of Tnni3^+/+, Tnni3^R186Q/R186Q and Tnni3^R186Q/R186Q +C75 mice ventricular myocardium (cross-section). With HE staining (400×), blue represents the nucleus, and pink represents the cytoplasm. With Masson’s trichrome staining (400×), the collagen fiber is shown blue and the cardiomyocytes are red. Scale bar 100 µm. (F) Quantification of total fibrosis by Masson’s trichrome staining in Tnni3^+/+, Tnni3^R186Q/R186Q and Tnni3^R186Q/R186Q +C75 mice ventricular myocardium (n = 3 per group). (G-H) Representative images of WGA staining in Tnni3^+/+, Tnni3^R186Q/R186Q and Tnni3^R186Q/R186Q +C75 mice ventricular myocardium, showing the cardiac myocyte (CM) cross-sectional area. Scale bar 20 µm. Quantitative data of CM hypertrophy assessed by cross-sectional area for WGA staining in Tnni3^+/+, Tnni3^R186Q/R186Q and Tnni3^R186Q/R186Q +C75 mice ventricular myocardium (n = 3 per group with 100 CMs analyzed). (I) Western blot analysis of the hypertrophic markers MYH7, ANP and BNP in ventricular myocardium heart tissue of Tnni3^+/+,Tnni3^R186Q/R186Q and Tnni3^R186Q/R186Q +C75 mice, and a representative experiment is shown (n = 3 per group). (J) The Western blot of the hypertrophic markers MYH7, ANP and BNP protein levels is expressed as the means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. The statistical test was done with unpaired T-test. GAPDH was used as the loading control.