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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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Compositional imaging analysis of focal adhesions upon ROCK perturbations. (A) Pixels of the 4-color live cell imaging of 5 cells clustered according to the fractional intensities of zyxin, FAK, vinculin and paxillin. (B) Bar plots presenting the average fractional intensity for each component in each cluster, defining the compositional signatures. (C) Compositional images of a cell (top) and of the marked (dashed line) region within it magnified (bottom) before and after the first Y-27632 addition. Pixels are colored according to their compositional signatures color-code, as shown in (B). Scale bar, 10 μm. (D) Bar plots showing the mean of the fold change (14 minutes after perturbation/before perturbation) in the number of pixels (abundance) assigned to each compositional signature (n = 5 cells, error bars indicate standard error of the mean). (E) The compositional signatures clustered based on their abundance profile after the various perturbations. The heatmap shows the mean abundance of each composition at the indicated time points after ROCK perturbations (n = 5 cells). The columns of the heatmap are ordered according to the hierarchical clustering dendrogram shown on top.