Fig. 7: Defects in both ciliogenesis and blood flow block EndoMT.
From: Endocardial primary cilia and blood flow regulate EndoMT during endocardial cushion development
a, snRNA-seq schema. b, UMAP plot of e9.5 hearts (29,313 nuclei). c, UMAP plot of the endocardial cluster (4,291 nuclei) from b. d, Volcano plot of DEGs between WT and cilia KO (Ift20−/−) endocardium (1,340 and 922 nuclei, respectively); KLF4 targets are highlighted. e, Gene set enrichment for EndoMT progression in vEC/EndoMT clusters (1,496 nuclei). f, UMAP plot of vEC/EndoMT clusters (1,496 nuclei). g, Pseudotime analysis of vEC/EndoMT clusters using Slingshot on WT and heterozygous littermate controls. h, Pseudotime progression of vEC/EndoMT clusters using Slingshot for all three genotypes. i, Percentage of nuclei in each EndoMT pseudostage per genotype (Cilia WT: nuclei = 516, Cilia Het (Ift20+/−): nuclei = 532, Cilia KO (Ift20−/−): nuclei = 448). Statistically significant P values are indicated in red (Mid EndoMT: Cilia WT versus Cilia KO P = 0.003, Cilia Het versus Cilia KO P = 0.14; Late EndoMT: Cilia WT versus Cilia KO P = 0.028, Cilia Het versus Cilia KO P = 0.026) Statistics: NS, P > 0.05; *P ≤ 0.05; **P ≤ 0.01. The statistical test used was a two-tailed t test with Bonferroni correction. Data are represented as mean ± s.e.m. Box plot statistics for i: vEC Cilia WT (min: 4.17, Q1: 5.87, median: 7.57, Q3: 9.11, max: 10.64, lower whisker: 4.17, upper whisker: 10.64), vEC Cilia Het (min: 9.01, Q1: 9.87, median: 10.72, Q3: 11.57, max: 12.43, lower whisker: 9.01, upper whisker: 12.43), vEC Cilia KO (min: 16.33, Q1: 21.30, median: 26.27, Q3: 31.24, max: 36.21, lower whisker: 16.33, upper whisker: 36.21), Early EndoMT Cilia WT (min: 42.11, Q1: 43.97, median: 45.83, Q3: 47.91, max: 50.00, lower whisker: 42.11, upper whisker: 50.00), Early EndoMT Cilia Het (min: 35.59, Q1: 38.03, median: 40.47, Q3: 42.91, max: 45.35, lower whisker: 35.59, upper whisker: 45.35), Early EndoMT Cilia KO (min: 14.95, Q1: 19.88, median: 24.82, Q3: 29.75, max: 34.69, lower whisker: 14.95, upper whisker: 34.69), Late EndoMT Cilia WT (min: 18.62, Q1: 23.90, median: 29.17, Q3: 31.86, max: 34.54, lower whisker: 18.62, upper whisker: 34.54), Late EndoMT Cilia Het (min: 23.73, Q1: 24.98, median: 26.23, Q3: 27.48, max: 28.73, lower whisker: 23.73, upper whisker: 28.73), Late EndoMT Cilia KO (min: 2.66, Q1: 3.53, median: 4.39, Q3: 5.25, max: 6.12, lower whisker: 2.66, upper whisker: 6.12), Mid EndoMT Cilia WT (min: 15.79, Q1: 18.27, median: 20.74, Q3: 20.78, max: 20.83, lower whisker: 15.79, upper whisker: 20.83), Mid EndoMT Cilia Het (min: 16.90, Q1: 19.74, median: 22.57, Q3: 25.41, max: 28.25, lower whisker: 16.90, upper whisker: 28.25), Mid EndoMT Cilia KO (min: 42.86, Q1: 43.69, median: 44.52, Q3: 45.35, max: 46.18, lower whisker: 42.86, upper whisker: 46.18). EC, endocardial cell; FACS, fluorescence-activated cell sorting; HE, hematoendothelium; PIM, proinflammatory endocardium; Prolif., proliferating; Het, heterozygous; FC, fold change; Endo, EndoMT; Mes, mesenchymal.
