Extended Data Fig. 8: Regulation of the potential ephrin-B2 receptor EPHA4.
From: A human cell atlas of the pressure-induced hypertrophic heart
(a) FeaturePlot for the expression of EPHA4 in snRNA-seq dataset (log2 transformed and normalized UMI counts). Color refers to the expression. (b) Linear regression for EPHB1 protein expression in cardiomyocytes (see Fig. 4i) and maximal wall thickness in mm (see Supplementary Data 3), shown for individual patients (Healthy n=14; AS n=5). The confidence bands reflect the standard error of the beta coefficients. (c) Violin plot for EPHA4 in hypertrophied (AS) vs. healthy cardiomyocytes (log2 transformed and normalized UMI counts). (d) Box plot showing expression of EPHA4 in cardiomyocytes in individual patients of respective groups (n=14 healthy, n=5 AS) (log2 transformed and normalized UMI counts; visualized as median and 25th and 75th percentiles with whiskers indicating maximal and minimal values). (e) Violin plot for EFNB2 in hypertrophied (AS) vs. healthy cardiomyocytes (log2 transformed and normalized UMI counts). (f) Phosphorylated EPHA4 protein in human cardiomyocytes treated with recombinant ephrin-B2 (10μg/ml, 15min), compared to non-treated CMs (=Ctr). Left: Quantification displayed as fold change vs Ctr (n=3; normalized to α-Tubulin-1B); Right: representative Western blot. α-Tubulin-1B served as loading control. Blots were processed in parallel and the loading control run on the same blot. Adjusted p-values based on Bonferroni correction using all genes of the dataset to compare the expression in violin plots were calculated with the Seurat function ‘FindAllMarkers’ using ‘bimod’ as the statistical test (c, e). Data are shown as mean ± s.e.m. (f). Normal distribution was assessed using the Kolmogorov-Smirnov test (d, f). t=0,05916, four degrees of freedom (f). For Gaussian distributed data, statistical analysis to compare two groups was performed using the unpaired, two-sided Student’s t-test (d, f).
