Fig. 5
Endothelin-1 induces iPericyte contraction. A, B Normalised cell index of neural crest iPericytes, mesoderm iPericytes and HBVPs treated with endothelin-1 or vehicle (CPM) over a period of 2 h (n = 4 per condition). C–E Quantified AUC (C; indicator of volume of contraction), ∆ cell index (D; maximum contraction) and ∆ cell index after 2 h (E; contraction at 2 h time point) for mesoderm iPericytes and HBVPs treated with control or endothelin-1 analysed using two-way ANOVA: AUC (cell type: F (1, 12) = 0.6953, p = 0.4206; treatment: (F (1, 12) = 13.35, p = 0.0033; interaction: F (1, 12) = 0.1006, p = 0.7565); ∆ cell index (cell type: F (1, 12) = 0.02309, p = 0.8817; treatment: F (1, 12) = 15.21, p = 0.0021; interaction: F (1, 12) = 0.5773, p = 0.4620); ∆ cell index after 2 h (cell type: F (1, 12) = 1.590, p = 0.2313; treatment: F (1, 12) = 14.31, p = 0.0026; interaction: F (1, 12) = 0.5518, p = 0.4719). F–H Quantified AUC (F), ∆ cell index (G) and ∆ cell index after 2 h (H) for neural crest iPericytes and HBVPs treated with control or endothelin-1 analysed using two-way ANOVA: AUC (cell type: F (1, 12) = 1.563, p = 0.2351; treatment: (F (1, 12) = 54.67, p < 0.0001; interaction: F (1, 12) = 5.470, p = 0.0375); ∆ cell index (cell type: F (1, 12) = 13.53, p = 0.0032; treatment: F (1, 12) = 66.11, p < 0.0001; interaction: F (1, 12) = 18.47, p = 0.0010); ∆ cell index after 2 h (cell type: F (1, 12) = 34.64, p < 0.0001; treatment: F (1, 12) = 38.56, p < 0.0001; interaction F (1, 12) = 14.70, p = 0.0024). C–H Post-hoc comparisons performed using Sidak’s multiple comparisons test. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. Data shown as mean ± SD. I–J Normalised gene expression counts of differentially expressed endothelin-1 receptors in HBVP, neural crest iPericytes and mesoderm iPericytes compared using DEseq: HBVPs and neural crest iPericytes EDNRA, I log2FoldChange = 2.53, padj = 6.12E−23; EDNRB, J log2FoldChange = 5.14, padj = 9.13E−26; neural crest iPericytes compared to mesoderm iPericytes EDNRA, I log2FoldChange = − 2.52, padj = 3.12387E−25; EDNRB, J log2FoldChange = − 7.22, padj = 6.77016E−15