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. 2019 Aug 27;9:12421. doi: 10.1038/s41598-019-48741-w

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© The Author(s) 2019

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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Confocal immunofluorescence microscopy. Adherent LPM from wild-type mice were dual labeled with FITC-phalloidin and anti-SerpinB2 antibody. (a) Actin; FITC-phalloidin (F-actin) staining showing actin concentrations at focal adhesions (green). SerpinB2; SerpinB2 staining localized around and in the focal adhesions (red). Merge; overlapping staining (yellow); see Fig. S5 for enlargements and quantitation of overlap. (b) Actin; FITC-phalloidin staining showing F-actin concentration along the leading edge of a lamellipodia (bottom left, green). SerpinB2; SerpinB2 staining (red) also staining the leading edge of a lamellipodia (bottom left, red). Merge; overlapping staining at the leading edge of a lamellipodia (yellow, arrowheads). (c) The two clusters of focal adhesions indicated in b (arrows) are enlarged to show overlapping localization of actin and SerpinB2 staining. The images shown are representative of 2 independent experiments where 5 images were examined containing ≈10 cells per image with ≈50 cells examined per experiment. Approximately 30% of cells examined showed overlapping staining at focal adhesions and/or lamellipodia.