Fig. 3: EPHB1 is dysregulated in cardiomyocytes of hypertrophied hearts.

a, Mean expression (log₂-transformed and normalized UMI counts) of Eph receptors in all cardiomyocytes. b, Heatmap of Eph receptors expressed (log₂-transformed and normalized UMI counts) in all cardiomyocytes of healthy individuals versus patients with AS. Colors indicate expression levels in each nucleus. c, Differential expression of Eph receptors in cardiomyocytes between hearts of healthy individuals versus patients with AS, ranked by differential expression and calculated by subtracting mean expression of genes (normalized UMI counts; values from individuals with hypertrophy minus those from healthy individuals). d, FeaturePlot for expression of EPHB1 (log₂-transformed and normalized UMI counts). Color reflects expression levels in each nucleus. e, EPHB1 expression (log₂-transformed and normalized UMI counts) according to cell type. f, Violin plot for EPHB1 expression in cardiomyocytes (log₂-transformed and normalized UMI counts). g, Violin plot for EPHB1 expression in cardiomyocytes from patients with AS versus healthy individuals shown for individual patients (log₂-transformed and normalized UMI counts). h, Box plot showing expression of EPHB1 in cardiomyocytes in individual patients (healthy, n = 14; AS, n = 5; log₂-transformed and normalized UMI counts, visualized as median with 25th and 75th percentiles, with whiskers indicating maximal and minimal values). i, Left, representative immunofluorescence images of cryosections from non-hypertrophied hearts and hearts from patients with hypertrophic cardiomyopathy. Blue, 4,6-diamidino-2-phenylindole (DAPI); red, EPHB1. Scale bar, 50 µm. Right, quantification for histological assessment of EPHB1 protein expression in n = 4 non-hypertrophied versus n = 5 hypertrophied hearts. FC, fold change. j, Left, representative immunofluorescence images of cardiac cryosections from sham-operated mice and operated mice in the TAC model. Blue, DAPI; red, EPHB1. Scale bar, 50 µm. Right, quantification for histological assessment of EPHB1 protein expression in n = 4 hearts of sham-operated mice versus n = 6 hearts of mice in the TAC model. Adjusted P values based on Bonferroni correction using all genes in the dataset to compare expression in violin plots were calculated with the Seurat function ‘FindAllMarkers’ using ‘bimod’ as the statistical test (f,g). Data are shown as mean ± s.e.m. (i,j). Normal distribution was assessed using the Kolmogorov–Smirnov test (h–j). Statistical analysis to compare two groups was performed using unpaired, two-sided Student’s t-tests (h–j). t = 4,118, seven degrees of freedom (i); t = 2,477, eight degrees of freedom (j).

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