Vangl function in murine telocytes is required for the formation of Wnt deficient intestinal crypt.A Telocytes stained with FITC-phalloidin. Yellow arrows indicate filopodia protrusions. Blue arrows indicate thicker protrusions. Scale bar = 10 µm. B, C Number of filopodia per cell and average length of filopodia in telocytes treated with control scrambled siRNA and siRNA for Vangl1, Vangl2 and Vangl1/Vangl2, IRSp53. B (n = 35, 21, 22, 27, 6 cells). C (n = 61, 50, 30, 30, 6 cells, n = 3488, 1796, 1964, 1079, 92 filopodia). Graphs represent mean and SEM. Statistical significance: p ≤ 0.05. B, C Two-sided Kruskal-Wallis tests with Bonferroni correction for multiple tests. SEM = 1. D Organoid formation assay: PDGFRα-cre/Rosa2-mTmG telocytes (green), co-cultured with Porcn−/−- intestinal epithelial crypt cells (brightfield) in scrambled siRNA, Vangl1, Vangl2, Vangl1/Vangl2, and IRSP53 siRNA treated conditions. Micrographs were taken day 1 after co-culture, indicating that cells targeted with control or specific siRNAs were establishing extensive contacts with organoid epithelial cells. Arrows indicate long filopodia-like membrane protrusions in siRNA treated control group while in groups treated with Vangl1/2 or IRSp53 siRNA contacts are established via lamellipodia and thicker, telome-like structures. Scale bar = 10 µm. E Organoid formation assay of Porcn deficient crypt cells co-cultured with murine telocytes treated with control scrambled, Vangl1, Vangl2, or Vangl1/Vangl2 siRNA. Organoid counts were recorded per condition (n = 4 assays per condition). F Organoid formation assay of Porcn deficient crypt cells co-cultured with murine telocytes treated with control scrambled, IRSP53 siRNA. (n = 4 assays per condition). Source data are provided as a Source Data file.
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