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. 2018 Nov 27;9:5021. doi: 10.1038/s41467-018-07448-8

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© The Author(s) 2018

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 — Regulation of Vinculin localization. a Colocalization between E-cadherin and Vinculin. Various E-cadherin clusters (arrowheads) colocalize with those of Vinculin. b Top panels, zoom-in view of boxed junction in a. Bottom panel shows the similarities in the intensity profiles for Vinculin and E-cadherin. c Representative images showing the distribution of Vinculin in water-injected embryo (left) and α-Catenin dsRNA injected embryo (right). d, e Quantifications showing a reduction in Vinculin recruitment and a loss of the planar polarized distribution of Vinculin due to α-Catenin RNAi. f–q Rok inhibitor H1152 was injected @ 20 mM concentration to inhibit Myosin-II activity. f, i, l, o Representative images showing the distribution of Myosin-II, Vinculin, E-cadherin, and Vinc/E-cad ratio, respectively, in water-injected control embryos (left panels) and H1152-injected embryos (right panels). g, h Quantifications showing a reduction in junctional Myosin-II recruitment and a loss of its planar polarized distribution due to Rok inhibition. i, k Quantifications showing a reduction in Vinculin recruitment and an inversion of its planar polarized distribution due to Rok inhibition. m, n Quantifications showing the reduction in E-cadherin levels and an amplification of its planar polarized distribution due to Rok inhibition. Corresponding representative images and quantifications for changes in Myosin-II distribution are presented in Figure 4a, c, d. p, q Quantifications showing a reduction in the Vinc/E-cad ratio and a loss of its planar polarized distribution due to Rok inhibition. Scale bar in b is 1 μm. All other scale bars represent 5 μm. All error bars represent SEM. Statistical significance estimated using two-tailed Student’s t-test. Images/quantifications in a–e and o–q come from embryos co-expressing Vinculin-mCherry and E-cadherin-GFP. Junctions marked based on E-cadherin localization. Images/quantifications in f–k come from embryos co-expressing Vinculin-mCherry and MyoII-GFP. Junctions marked based on Vinculin localization. Images/quantifications in l–n come from embryos co-expressing MyoII-mCherry and E-cadherin-GFP. Junctions marked based on E-cadherin localization. Insets in e, h, k, n and q indicate number of embryos. ns, p>0.05; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p<0.0001