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. 2021 Dec 10;12:7186. doi: 10.1038/s41467-021-27319-z

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — A E14.5 intestines after separation from the mesentery of Cilk1^+/− and Cilk1^−/− embryos. The length of the small intestine was measured from the caudal stomach (orange arrowheads) to the rostral cecum (purple arrowheads). Scale bar, 1 mm. B Lengths of the small intestines of Cilk1^+/+ (n = 9, 4, 5, 5, 4), Cilk1^+/− (n = 18, 7, 18, 12, 9), and Cilk1^−/− (n = 10, 5, 9, 7, 7) embryos at E13.5, 14.5, 15.5, 16.5, and 17.5. C E16.5 esophagi of Cilk1^+/− and Cilk1^−/− embryos. The length of the esophagi was measured from the stomach (orange arrowheads) to the pharynx (purple arrowheads). Scale bar, 1 mm. D Lengths of the esophagi of Cilk1^+/+ (n = 4 and 4), Cilk1^+/− (n = 4 and 8), and Cilk1^−/− (n = 5 and 4) embryos at E14.5 and E16.5 (adjusted P values are 0.0021 for Cilk1^+/+ vs Cilk1^−/− and 0.0003 for Cilk1^+/− vs Cilk1^−/−). E Immunofluorescence staining of E13.5 intestinal cross sections from Cilk1^+/− and Cilk1^−/− embryos for IFT88 (green), ciliary axonemes (TUB^Ac, red), basal bodies (γ-tubulin, white), and nuclei (Hoechst, blue). Right, higher magnifications of boxed cilia. White boxes highlight mesenchymal cilia and gray boxes highlight epithelial cilia. Scale bar of larger images, 5 μm. Scale bar of enlarged images, 2 μm. F Lengths of E13.5, E15.5, and E18.5 control (Cilk1^lox/lox n = 3, 5, 3 and Dermo1^Cre Cilk1^lox/+ n = 5, 7, 6) and Dermo1^Cre Cilk1^lox/lox (n = 6, 10, 4) small intestines. G Immunofluorescence staining for EdU (white) and nuclei (Hoechst, blue) in E13.5 Dermo1^Cre Cilk1^lox/+ and Dermo1^Cre Cilk1^lox/lox intestines. Scale bar, 25 μm. Orange dotted lines outline the epithelium. H The percentage of EdU-positive cells in the whole intestine, the epithelial cells and the mesenchymal cells in Cilk1^lox/lox (n = 3), Dermo1^Cre Cilk1^lox/+ (n = 4) and Dermo1^Cre Cilk1^lox/lox (n = 4) embryos at E13.5. For each intestine, we averaged the percentages from three sections. B, D, F, and H Each point in the scatter plots represents the value from an individual embryonic intestine or esophagus. Horizontal bars indicate means ± SD. ns, p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001 and ****p < 0.0001 indicate adjusted P values between indicated samples by two-way ANOVA Tukey’s multiple comparisons test. Source data are provided in the Source Data file.