Fig. 6: Regionally restricted sex differences in vascular smooth muscle.
From: Region-specific gene expression and sex inform about disease susceptibility in the aorta
a, Schematic of experimental design. Twelve-week-old C57BL/6J female and male mice (four per group or sex) were dissected and digested into eight single cells for generation of libraries based on location and sex for scRNA-seq. b, UMAP of vSMCs from the eight libraries. c, Heatmap of all 527 sex-enriched transcripts in vSMCs. Dashed boxes indicate concurrent gene expression patterns across sex rather than anatomic site. d, Number of sex-enriched genes by anatomic location and sex (see Extended Data Fig. 2 for additional specifics). e, Gene ontology plot of uniquely enriched, autosomal thoracic-derived vSMC genes in both sexes calculated via hypergeometric distribution. P values for enrichment categories are as follows: female: electron transport chain, 9.82 × 10−8; establishment of cell polarity, 7.19 × 10−7; axon guidance, 7.94 × 10−7; male: peptide metabolic process, 6.15 × 10−12; negative regulation of catalytic activity, 1.80 × 10−10; extracellular matrix organization, 1.11 × 10−7. f, Representative IF images of BGN expression (green) in female and male thoracic vSMCs. g, Quantification of BGN intensity in female and male vSMCs. Females, n = 54 images, 8 animals; males, n = 21 images, 4 males; 74 images in total. P < 0.0001. h, Violin plot visualizing the expression pattern of Bgn in thoracic vSMCs by sex. i, Representative IF images of POSTN expression (green) in female and male thoracic vSMCs. j, Quantification of POSTN intensity in female and male vSMCs. Females, n = 31 images, 5 animals; males, n = 18 images, 5 males; 49 images in total. P < 0.0001. k, Violin plot visualizing the expression pattern of Postn in thoracic vSMCs by sex. l, Gene ontology plot of uniquely enriched, autosomal abdominal vSMC genes in both sexes calculated via hypergeometric distribution. P values for enrichment categories are as follows: females: diabetic cardiomyopathy, 2.27 × 10−22; ATP biosynthetic process, 1.99 × 10−10; modulation of host immune response, 1.99 × 10−7; males: NABA core matrisome, 4.27 × 10−7; myometrial contraction, 2.60 × 10−6; response to growth factor, 3.42 × 10−6. m, Representative IF images of TGFBI expression (green) in female and male abdominal vSMCs. n, Quantification of TGFBI intensity in female and male vSMCs. Females, n = 21 images, 5 animals; males, n = 14 images, 3 males; 35 images in total; P < 0.0001. o, Violin plot visualizing the expression pattern of Tgfbi in abdominal vSMCs by sex. p, Representative IF images of MCAM expression (green) in female and male abdominal vSMCs. q, Quantification of MCAM intensity in female and male vSMCs. Females, n = 30 images, 8 animals; males, n = 26 images, 7 males; 56 images in total; P < 0.0001. r, Violin plot visualizing the expression pattern of Mcam in abdominal vSMCs by sex. For g,j,n,q, error bars represent mean ± s.d. n = 14–25 images per animal (gray), 3–8 animals per sex (blue squares, male; pink inverted triangles, female). Two-sided Mann–Whitney test (Postn, Bgn, Mcam) and two-sided Welch’s t-test (Tgfbi). Panel a created using BioRender.com.
