Extended Data Fig. 6: JAG1-NOTCH2 Mediated Notch Signal Between UDVECs and Elastin-VICs.
From: APOE–NOTCH axis governs elastogenesis during human cardiac valve remodeling
a. Major Extracellular Matrix (ECM)-related Ligand-Receptor (L-R) pairs between Elastin-VICs and unidirectional VECs (UDVECs) subtypes; b. Total L-R pair numbers between UDVECs and Elastin-VICs; c. Notch-related signaling patterns from UDVECs to Elastin-VICs; d. Gene Ontology (GO) enrichment analysis of differential expressed genes within each VIC subtype; e. Violin plot of DLL1 in VECs. FDR: PTGDS-VECs vs. other VEC subtypes; f. RNA in situ hybridization staining of PTGDS, JAG1, and NOTCH3 in four human fetal valves at W15, n = 3 different hearts; g. Violin plots of NOTCH1-3, HES1, HES4, and HEY2 in VICs. FDR: Elastin-VICs vs. other VIC subtypes; h. Immunofluorescence staining of NOTCH3, NOTCH1, HES1, and Elastin in four different valves; i. Violin plots of Notch receptors and downstream target gene expressions within Elastin-VICs (Left), and Notch ligands in UDVECs (Right) across each valve; j. qPCR analysis of Notch target genes in VICs with Jag1 (15 μg/ml) treatment; k. qPCR analysis of Notch receptors and target genes in cultured human VICs after combinational KD of NOTCH receptors. Jag1 (15ug/ml) was included; n = 3 biological repeats. Data shown as mean ± SEM. ns p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001. For g, i: Statistics adhered to a two-sided approach. Significance was determined by adjusted p-values (FDRs) < 0.05. For j, Control vs. Jag1 treatment. Unpaired 2-tailed t-test (two groups); P values for j: HEY2: p = 0.0006, HEYL: p = 0.0265, HES1: p = 0.2450. For k: one-way ANOVA followed by Tukey’s test, vs Scramble. P values for k: NOTCH2: Scramble vs. siN1: p > 0.9999, vs. siN2: p = 0.0008, vs. siN3: p = 0.1982, vs. siN1 + N2: p = 0.0024, vs. siN1 + N3: p > 0.9999, vs. siN2 + N3: p = 0.0008, vs. siN1 + N2 + N3: p = 0.0004, NOTCH1: Scramble vs. siN1: p < 0.0001, vs. siN2: p = 0.0025, vs. siN3: p = 0.0095, vs. siN1 + N2: p < 0.0001, vs. siN1 + N3: p < 0.0001, vs. siN2 + N3: p = 0.0206, vs. siN1 + N2 + N3: p < 0.0001, NOTCH3: Scramble vs. siN1: p = 0.3875, vs. siN2: p = 0.0053, vs. siN3: p < 0.0001, vs. siN1 + N2: p = 0.0038, vs. siN1 + N3: p < 0.0001, vs. siN2 + N3: p < 0.0001, vs. siN1 + N2 + N3: p < 0.0001, HEY2: Scramble vs. siN1: p = 0.6668, vs. siN2: p = 0.0413, vs. siN3: p = 0.9996, vs. siN1 + N2: p = 0.0510, vs. siN1 + N3: p = 0.9210, vs. siN2 + N3: p = 0.0238, vs. siN1 + N2 + N3: p = 0.0122, HEYL: Scramble vs. siN1: p = 0.2549, vs. siN2: p = 0.0001, vs. siN3: p = 0.0745, vs. siN1 + N2: p < 0.0001, vs. siN1 + N3: p = 0.0014, vs. siN2 + N3: p = 0.0001, vs. siN1 + N2 + N3: p < 0.0001. ECM: Extracellular Matrix, L-R: Ligand-Receptors.
