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. 2017 Jun 13;7:3414. doi: 10.1038/s41598-017-03640-w

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© The Author(s) 2017

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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FAK is activated through α2 integrins harboring the loss-of-function mutation E366A in the α2 subunit, and in the presence of EDTA. (a) SaOS cells with the loss-of-function mutation E336A in the α2 subunit (SaOS^α ^2E336A) on EV1-coated surface. Photographed through a phase contrast microscope after 0, 20, 40, and 60 min. Scale bar 50 μm. (b) Western blot analysis of pY397FAK in SaOS^α ^2E336A cells plated on EV1. Typical experiment on the left, and on the right, a visualization of the results from four independent experiments presented in box blot as natural logarithm of pY397FAK band intensity normalized to β-actin. Whiskers present minimum and maximum of all of the data. Non-parametric Mann-Whitney U test was used to determine statistical significance. (c) Phase contrast microscopy images of SaOS^α ²⁺ cells treated with integrin inactivating EDTA (5 mM) after spreading for 60 min on an immobilized layer of EV1 or collagen I. Scale bar 50 μm. (d) Western blot analysis of pY397FAK in SaOS^α ²⁺ and SaOS^α ^2E336A cells treated with 5 mM EDTA and plated on an immobilized layer of EV1.