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. 2019 Dec 27;9:19961. doi: 10.1038/s41598-019-56509-5

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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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Histological finding of reduced proliferation after treatment supports mRNA expression data. Paraffin-embedded treated and untreated peritumoral brain (a) and GBM tissue (b) was stained with a Ki67 antibody as proliferation marker (red) and DAPI as nuclei marker (blue) and recorded by a slide scanner. Representative images of untreated samples are presented. (Note: green signals are attributed to autofluorescence of the tissue). For quantification, the total tissue area (a’/”, b’/”, gray), the nuclei area (a’, b’) and the Ki67-positive nuclei area (a”, b”) were determined. H/E stainings of consecutive tissue sections are shown in a”’ and b”’. (c) Ratio of proliferating area (Ki67- and DAPI-positive pixel area) per DAPI area in untreated and treated (TMZ + 4 Gy) peritumoral brain (left) and GBM tissue (right). (d) Ratio of DAPI area per total tissue area compared to total tissue area in pixels in untreated and treated (TMZ + 4 Gy) peritumoral brain (red circles, green squares) and GBM tissue (blue diamonds, black triangles). Biological replicates: 1; Technical replicates: 3; Scanned sections: 33 (untreated peritumoral brain), 32 (treated peritumoral brain), 13 (untreated GBM), 8 (treated GBM). Scale bars: 100 µm (a), 50 µm (b).