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. 2018 Feb 1;8:2187. doi: 10.1038/s41598-018-20252-0

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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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Four-color live cell imaging of focal adhesions responding to ROCK perturbations. (A) A scheme indicating the four examined components and their vertical positions within the focal adhesion²⁸. Inhibition and recovery of actomyosin contractility were achieved by the addition and washout of the ROCK inhibitor Y-27632, respectively. (B) Images of a cell expressing TDmKate2-zyxin, mCitrine-FAK, TDmTFP-vinculin and TDmTagBFP-paxillin before the perturbations. Scale bar, 10 μm. (C) Heatmaps show the integrated density of zyxin, FAK, vinculin and paxillin in each of the tracked focal adhesions of one cell as a function of time. Corresponding rows in the four heatmaps relate to the same focal adhesion. Arrows indicate the time of Y-27632 (Y) additions and washouts (W). The anchor image was obtained from Wikimedia Commons.