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. 2018 Mar 21;8:4986. doi: 10.1038/s41598-018-22940-3

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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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Tether-mediated cell-cell adhesions are anchored via adherens-type junctions. (A,C) Densely-plated 4T1 cells form E-cadherin- (A) and β-catenin-rich (C) adherens junctions, located either along the sub-apical region (arrows), or in multiple dorsal patches (arrowheads). (B,D) Sparsely plated 4T1 cells form structurally and molecularly polarized tethers that are attached to the neighboring cell via E-cadherin (B), co-localized actin (magenta) and β-catenin (yellow) (D). Arrowhead in Panel D denotes the tether adhesion area. Scale bar: 10 μm. (E,E’) Immunoblotting and corresponding histogram depicting E-cadherin levels in sparsely- (5.6*10³/cm²) and densely-plated (8.9*10⁴/cm²) 4T1 cells. The normalized levels of E-cadherin (relative to α-tubulin) are shown in E’. Blotting Lanes of Fig. 3E were extracted from the same gel (full blotting of the gel is shown in Supplementary). (F) Comparison of E-cadherin mRNA levels in densely- and sparsely-plated 4T1 cells normalized to HPRT1 and GUSB genes. The results are based on two independent experiments.