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. 2018 Apr 27;9:1688. doi: 10.1038/s41467-018-04030-0

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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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Fig. 2 — Subcellular protrusion phenotypes revealed by a time series clustering analysis. a–c A representative cluster assignment on a time-lapse movie of a PtK1 cell stained with CellMask DeepRed. Edge evolution on 5 s interval (a), cluster assignments of each probing window on every four frame (20 s interval) (b), and the space-time maps of instantaneous edge velocity (c) of the entire cell edge and the indicated cluster regions. Scale bar: 10 μm. d Raw velocity maps for Cluster I, II-1, II-2, II-3, and III. All time series are registered with respect to protrusion onset (t = 0). e Average time series of protrusion velocity registered at protrusion onsets (t = 0) in each cluster. Solid lines indicate population averages. Shaded error bands indicate 95% confidence intervals of the mean computed by bootstrap sampling. n indicates the number of time series in each cluster. The time lapse movies of 36 cells were used in this analysis. f Proportions of each cluster in entire samples or individual cells expressing fluorescent actin, Arp3, VASP, and HaloTag, respectively. g Decision graph of the density peak clustering analysis of protrusion velocities. h A t-SNE plot of the autocorrelation functions of protrusion velocity time series overlaid with cluster assignments. i Spatial conditional distribution of each cluster. Solid lines indicate population averages. Shaded error bands indicate 95% confidence intervals of the mean computed by bootstrap sampling