Fig. 7: Sex as a modifier of the MCAM-associated transcriptional signature in aortic vSMCs.
From: Region-specific gene expression and sex inform about disease susceptibility in the aorta
a, Localization of MCAM in the thoracic–abdominal region in the mouse aorta. Sk. muscle, skeletal muscle. b, Protein lysates from a cohort of female and male vSMC cell lines were probed for expression of MCAM and GAPDH (loading control). c, Quantification of the relative ratio of MCAM to GAPDH in each donor. Data are shown as mean ± s.e.m; n = 3 (female), n = 6 (male); two-sided Welch’s t-test, P = 0.0173. d, Experimental design. Aortic vSMCs isolated from three female and three male donors were cultured and transfected with siRNA against MCAM. Forty hours after transfection, RNA was isolated for transcriptomic profiling. KD, knockdown. e, Heatmap visualizing transcriptional expression of selected markers of biological sex and vSMC identity across experimental conditions and donors. f, Venn diagram visualizing the overlap of differentially expressed genes (DEGs) in male versus female vSMCs following knockdown of MCAM. g–i, Gene ontology enrichment for sex-invariant (g), male-enriched (h) and female-enriched (i) differentially expressed genes in MCAM-knockdown vSMCs, 40 h after transfection. Dot color indicates the direction of expression change upon loss of MCAM expression, while size indicates significance of enrichment; P values are as follows: sex invariant (g): cell cycle, P = 5.18 × 10−78; DNA metabolism, P = 6.65 × 10−55; DNA replication, P = 1.84 × 10−33; male enriched (h): cytokine signaling, P = 4.11 × 10−10; cell migration, P = 7.47 × 10−10; cholesterol synthesis, P = 2.19 × 10−9; female enriched (i): VEGF signaling, P = 4.30 × 10−12; intracellular signal transduction, P = 4.49 × 10−11; vascular development, P = 1.09 × 10−10. For g–i, selected graphs of transcript levels of member genes in each ontology category data are visualized using mean ± s.e.m. n = 6. Displayed P values are Benjamini–Hochberg corrected Wald test values from DESeq2 for the following genes: BRCA2 (male, P = 1.29 × 10−17; female, P = 7.56 × 10−37), TK1 (male, P = 9.39 × 10−26; female, P = 3.62 × 10−37), CENPE (male, P = 2.15 × 10−40; female, P = 1.34 × 10−20), SREBF2 (male, P = 2.18 × 10−13; female, P = 9.84 × 10−9), PTGS2 (male, P = 3.01 × 10−76; female, P = 0.62), CX3CL1 (male, P = 1.14 × 10−19; female, P = 5.11 × 10−6), MYLK (male, P = 0.50; female, P = 9.85 × 10−11), GDF15 (male, P = 0.04; female, P = 6.81 × 10−30), F3 (male, P = 0.34; female, P = 3.79 × 10−47). Mouse pictograph (a), human pictograph (b) and panel d created using BioRender.com.
