Fig. 6: Failure of ciliogenesis results in abnormal KLF4 expression.
From: Endocardial primary cilia and blood flow regulate EndoMT during endocardial cushion development
a, KLF4 protein-positive endocardial cells in the AVC over time as a percentage of all endocardial cells in WT mice (blue) and cilia KO mice (green) (e8.5 (WT n = 6, cilia KO n = 6), e9.0 (WT n = 3, cilia KO n = 4), e9.5 (WT n = 6, cilia KO n = 6)) (e8.5 P = 0.0219, e9.0 P < 0.0001, e9.5 P = 0.0003). b, Immunofluorescence on e9.5 WT and cilia KO (Kif3a−/−) whole-mount hearts for KLF4 (red) in endocardial cells (CD31, white). Nuclei are shown in blue (Hoechst); the AVC is outlined in white. c, KLF4 protein-positive endocardial cells versus distance in e9.5 AVCs of cilia KO mice (green, n = 3). For comparison, WT is shown in blue (n = 4). d, Klf4 mRNA expression measured by the gray value over distance in the AVC of e9.5 WT mice (blue, n = 3) and cilia KO mice (green, n = 4). Shading indicates ±s.e.m. from the mean (line). e, HCR-FISH on whole-mount WT and cilia KO mouse embryos at e9.5 showing Klf4 mRNA (red) and nuclei (Hoechst, blue). Endocardial regions are outlined in white. f, KLF4 protein-positive endocardial cells in the AVC and OFT as a percentage of all endocardial cells in e9.5 WT (blue) and Dnah11−/− (gray) mice (AVC: WT n = 6, Dnah11−/− n = 6; OFT: WT n = 6, Dnah11−/− n = 6). g, Immunofluorescence on e9.5 sections of Ift20+/−/Ncx1+/− and Ift20−/−/Ncx1−/− hearts for KLF4 (red) in endocardial cells (CD31, white). Nuclei are shown in blue (Hoechst); the AVC is outlined in white. h, KLF4 protein-positive endocardial cells in the e9.5 AVC as a percentage of all endocardial cells in Ift2+/−/Ncx1+/− (blue, n = 3) and Ift20−/−/Ncx1−/− (brown, n = 4). Ift20−/− (green, n = 4) and Ncx1−/− (red, n = 4) are provided for comparison (Ift20+/−/Ncx1+/− versus Ift20−/− P = 0.0192, Ift20+/−/Ncx1+/− versus Ncx1−/− P = 0.0043, Ift20+/−/Ncx1+/− versus Ift20−/−/Ncx1−/− P = 0.0119, Ift20−/−/Ncx1−/− versus Ncx1−/− P = 0.6142). For c and d, distance runs from left to right: atrium to left ventricle. Statistics: NS, P > 0.05; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001. The statistical test used was a two-sided t test with Welch’s correction. Data are represented as mean ± s.e.m. Unless otherwise stated, n is the number of embryos. DA, dorsal aorta.
