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. 2019 Jun 20;2:226. doi: 10.1038/s42003-019-0462-y

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© The Author(s) 2019

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. 4 — Appearance and motility of nodal cilia in Efcab1^−/− mice. a Scanning electron microscopy of E7.5 embryo and nodal cilia in Efcab1^+/+ and Efcab1^−/− mice. Left, whole embryo; middle, node area; right, magnified image of nodal cilia. b Number of motile cilia in the node. N (embryos) = 18 (Efcab1^+/+) and N = 18 (Efcab1^−/−). c, d Nodal flow in Efcab1^+/+ (c) or Efcab1^−/− (d) E7.5 embryos. Upper; Trajectories of fluorescent beads driven by nodal flow. Sixty images acquired at 0.25 s intervals are superimposed. Red and blue dots indicate the start and end points of the superimposition, respectively. Lower; direction and distance traveled by fluorescent beads in Efcab1^+/+ or Efcab1^−/− nodal flows. Arrow length represents bead path and distance for 5 s. A, anterior; P, posterior; R, right; L, left. e Flow velocity of fluorescent beads. Boxes correspond to the first and third quartiles, and whiskers extend to the full range of the data. N (beads) = 112 from four embryos (Efcab1^+/+) and N = 46 from 10 embryos (Efcab1^−/−). f, g Trajectories of fluorescent beads binding to the tip of nodal cilia in Efcab1^+/+ (f) and Efcab1^−/− (g) mice. h Whole-mount in situ hybridization analysis of Nodal and Lefty expression in E8.0 embryos. R, right; L, left. Scale bars: 200 µm (a, left), 10 µm (a, middle, c, d), 1 µm (a, right), 4 µm (f, g), 1 mm (h). i Immunofluorescent localization of Efcab1 in the central and peripheral regions of node in Efcab1^+/+ and Efcab1^−/− mice. Images with anti-Efcab1 (green) and anti-acetylated-α-tubulin (red) antibodies are shown. Scale bars: 10 µm. **p < 0.01, ***p < 0.001 vs. Efcab1^+/+ (Student’s t-test)