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. 2019 Mar 21;9:5012. doi: 10.1038/s41598-019-41380-1

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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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Roscovitine (RSV) alone or in combination with Temozolomide (TMZ) restricts glioma progression in vivo. (A) H and E staining of tumor sections on 7^th day after tumor implantation was done in 2 randomly selected rats to confirm the development of tumor. Sections are shown at different magnification to show the spread of tumor from the site of injection (black arrow). (B) H and E staining of tumor sections (5–10 µ) from vehicle, TMZ, RSV and TMZ + RSV (from top to bottom) treated rats. After completion of the respective treatment cycles, rats were sacrificed. Brains were harvested and fixed in 4% PFA followed by sucrose gradient dehydration and cryosectioning. The sections thus obtained were used for H and E staining. Images were captured at 10 × and 4 × . Black arrows show the extensive presence of hemorrhages in vehicle treated tumors whereas hemorrhages were totally absent in all the treated tumors. (C) 25–30 µ sections were used for Immunohistochemical staining for Ki67 in tumor sections from vehicle, TMZ, RSV and TMZ + RSV treated rats. Black arrow shows clear nuclear staining for Ki67 in tumors from vehicle treated rats. Representative images of tumor sections are shown for each group.