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. 2018 Jul 30;9:2980. doi: 10.1038/s41467-018-05367-2

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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. 3 — SCABs suppress leading edge motility in a myosin contractility-dependent manner. a Leading edge kymography in MCF7 cells (top row) and T47D cells (bottom row) expressing iRFP-Lifeact (black) and MLC-mRuby2 (magenta) before and after dissolving SCABs using 25 µM ROCK inhibitor. Left panels are images from TIRF microscopy time-lapse series before and after the addition of ROCK inhibitor (Supplementary Movie 5). Scale bar is 10 µm. Line shows the leading edge position at which kymographs were registered. Right panel shows MLC kymograph (denoting SCABs) superimposed over Lifeact kymograph (demarcating the leading edge). Vertical scale bar is 10 µm and horizontal scale bar is 10 min. b Immunofluorescence of SCAB in MDCK epithelial cells and Caco-2 human intestinal epithelial cells. Representative images for F-actin, pMLC, and Arp2/3. Scale bar is 10 µm. c Leading edge kymography in MDCK cells (top row) and Caco-2 (bottom row) cells expressing iRFP-Lifeact (black) and MLC-mRuby2 (magenta) before and after dissolving SCABs using 25 µM ROCK inhibitor. Scale bar is 10 µm. Line shows the leading edge location at which the kymograph was registered. Right panel shows MLC kymograph (denoting SCABs) superimposed over Lifeact kymograph (demarcating the leading edge). Vertical scale bar is 10 µm and horizontal scale bar is 10 min