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. 2017 Aug 9;7:7700. doi: 10.1038/s41598-017-07043-9

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© The Author(s) 2017

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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IHC analyses of s.c. melanoma allografts stromal features. (a–f) Microscopic views of tissue sections IHC allowing visualization of intratumoral (a) TSP-1 (×20, black arrowheads), (c) CD3 (×100) and (e) VEGF (×20, dotted lines) immunostainings and quantification of (b) percentage of TSP-1-positive blood vessels as well as relative (d) CD3-positive and (f) VEGF-positive areas (mean ± SEM, t test, ns not significant, *p < 0.05, **p < 0.01, ***p < 0.001). All quantitative analyses were performed using ImageJ software. Five random fields were considered for TSP-1 and CD3 immunostainings, while the whole tumor section was analyzed for VEGF positivity.