Fig. 3: Mitochondrial and diastolic function in gonadectomized mice.

A, B Heart mtDNA copy number in C57BL/6 J male (A) and female (B) mice subjected to sham, gonadectomy or hormone replacement after gonadectomy under chow diet. mtDNAs were normalized to nuclear DNA. N = 4. A p = 0.019 (Sham/GDX) and 0.013 (GDX/Testosterone). B p = 0.036 (Sham/GDX), and 0.04 (GDX/Estrogen). C Experimental timeline for the gonadectomy and induction of HFpEF in C57BL/6 J male and female mice. D–I Oxygen consumption rates (OCR) of isolated mitochondria from C57BL/6 J male hearts were measured by Seahorse assay. The coupling assay measures basal respiration in presence of palmitoyl-carnitine, state 3 (ADP) and state 3 u (FCCP). The electron flow measures complex I (palmitoyl-carnitine), complex II (succinate), and complex IV (TMPD) respiration. N = 7. E p = 0.012. F p = 0.001. G p = 0.008. H p = 0.001. I p = 0.017. J–O Same Seahorse assays were performed in C57BL/6 J female mice after gonadectomy and 7 weeks of HFD + l-NAME feeding. Sham, n = 5; GDX, n = 6. K p = 0.013. L p = 0.006. M p = 0.049. P, Q E/e’ ratio (P) and LVEF (Q) of C57BL/6 J male mice after gonadectomy and 7 weeks of HFD + l-NAME diet. N = 11. P p = 0.027. R–T E/e’ ratio (R), E/A ratio (S), and LVEF (T) of C57BL/6 J female mice after gonadectomy and 7 weeks of HFD + l-NAME diet. Sham, n = 6; GDX, n = 7. R p = 0.0001. S p = 0.016. Each point represents a mouse. All data are presented as the mean ± SEM. ns not significant, *P < 0.05, **P < 0.01, and ***P < 0.001, by one-way ANOVA (A, B) or by two-sided Student’s t-test (D–T). Source data are provided as a Source Data file.