Fig. 5: Identification of Acsl6 as a sex-biased cis-regulator of diastolic dysfunction.

A Principal component analysis (PCA) plot of male and female C57BL/6 J mice under chow diet and 7 weeks of HFD + l-NAME diet. DEGs were identified with a false discovery rate (FDR) < 0.05. B Acsl6 mRNA levels in the hearts of male (n = 96) and female (n = 87) HMDP mice. p < 0.0001. C, D qRT-PCR (C) and western blotting (D) showing Acsl6 mRNA and protein in C57BL/6 J male and female hearts under chow and HFpEF condition. mRNA and protein levels were normalized to actin. N = 8. C p (male-chow/female-chow) < 0.0001; p (male-HFpEF/female-HFpEF) < 0.0001; p (female-chow/female-HFpEF) = 0.014. E qRT-PCR showing mRNA levels of Acsl6 in C57BL/6 J male and female heart after gonadectomy and 7 weeks of HFD + l-NAME diet. mRNA levels were normalized to actin. Male, n = 8; female, n = 6 (sham) or 7 (GDX). p (male) = 0.019; p (female) = 0.024. F Manhattan plot showing the significance (−log10 of p) of all SNPs and eQTL of Acsl6 in female mice after 3 weeks of isoproterenol infusion (female ISO-HMDP). The significant threshold (red line) of p = 4.1 × 10⁻⁶ is indicated. Association p-values from FaST-LMM are from Wald tests uncorrected for FDR. G, H E/A ratio distribution (G) and Acsl6 heart expression (H) based on genotype distribution at peak SNP associated with Acsl6 on chromosome 11 (rs26974045). Box and whisker plot depicting mean and distribution. Boxplots are shown centered on medians with upper and lower quartiles indicated by the box boundaries; whiskers enclose the box ± 1.5× the interquartile range (IQR). AA allele, n = 30; GG allele, n = 75. G p = 0.005. H p = 0.0002. I The correlation between local genetic variation in Acsl6 expression (cis-eQTL) and parameters of diastolic function and metabolism in ISO-HMDP. p-values are from biweight midcorrelation (bicor) tests. J, K Acsl6 expression (J) and E/e’ ratio (K) in male and female A/J and KK/HIJ strains of mice after 7 weeks of HFD + l-NAME feeding. J n = 4. K A/J-male, n = 7; A/J-female, n = 8; KK/HIJ-male, n = 6; KK/HIJ-female, n = 4. J p (A/J-male/A/J-female) = 0.025; p (A/J-male/KK/HIJ-male) < 0.0001; p (A/J-female/KK/HIJ-female) = 0.001; p (KK/HIJ-male/KK/HIJ-female) = 0.038. K, p (A/J-male/A/J-female) = 0.025; p (A/J-male/KK/HIJ-male) = 0.0001; p (A/J-female/KK/HIJ-female) < 0.0001; p (KK/HIJ-male/KK/HIJ-female) = 0.021. Each point represents a mouse. All data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****p < 0.0001, by two-way ANOVA (C, E, J, K) or by two-sided Student’s t-test (B, G, H). Source data are provided as a Source Data file.