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. 2018 Apr 18;8:6205. doi: 10.1038/s41598-018-24425-9

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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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Immunostaining for selected proteins encoded by up-regulated genes after injection of S100A8/A9 treated MSCs into wounds. Representative photomicrographs of sections from wounds which were injected with either control MSCs or S100A8/A9 treated MSCs. The expression of selected genes on protein level is shown with immunostaining for the serine protease PRSS36, the MSC specific chemokine receptor CCR7, and the developmental-like proteoglycan SPARC. Double staining (DP) for human β2 microglobulin (hβ2M is shown in green detecting the injected MSCs) and PRSS36 (shown in red) (a); CCR7 (shown in red) (b) and SPARC (shown in red) was compared to single staining with hβ2M. Nuclei were counterstained with DAPI. Scale bars: 200 μm. Dashed lines indicate the junction between epidermis (e) and dermis. es, eschar; wm, wound margin. Quantification of double-stained and single hβ2M⁺ MSCs in wounds injected with control MSCs and S100A8/A9 treated MSCs at days 2 after wounding were counted in 10 high-power fields per sample. Results are mean ± SD ratio of DP + and single hβ2M⁺ MSCs to total cells counted in the dermis (n = 6). ***P < 0.0001 versus non-treated control MSC injected wounds and MSCs pretreated with S100A8/A9 prior to be injected into wounds. The significance was obtained using ANOVA with Tukeys post hoc test.