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. 2020 Feb 7;10:2142. doi: 10.1038/s41598-020-58882-y

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© The Author(s) 2020

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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Vinculin is required for stable F-actin localization within uropods and neutrophil polarization. (a) Representative immunoflluorescence images of neutrophils after 30-minute incubation on immobilized ICAM-1 and CXCL1. Cells were stained with Hoescht, phalloidin, and anti-CD11a (n = 3 independent experiments). Scale bar = 10 µm. (b) Polarization of neutrophils based on asymmetric F-actin distribution (n = 3 independent experiments). (c,d) Background-subtracted and normalized TIRFM fluorescent intensities of F-actin and CD11a (n > 30 cells/group, 3 independent experiments). Analyzed using Kruskal-Wallis one-way ANOVA with Dunn’s multiple comparison test. ****p < 0.0001.