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. 2018 Oct 3;8:14770. doi: 10.1038/s41598-018-32878-1

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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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Poor perfusion instantly activated p53-mediated death signaling. In well-perfused brains (upper row), p53 immunolabeling (red) exhibits a predominantly cytoplasmic and diffuse staining (arrows, a), whereas anti-serine15-phospho-p53 antibodies weakly immunostained the nuclei (arrows, green, b). (c) Illustrates the intact nuclei identified by Hoechst staining (arrows, blue). The inset is the merged image of (a,b,c). In poorly perfused brain sections (lower row), p53 is still predominantly expressed in the cytoplasm but assumed a granular pattern (red, d). Triangles point at the swollen donut-like stained nuclei (d–f). These nuclei were not immunolabeled with phospho-p53 antibodies (green, e), whereas other intact nuclei preserved phospho-p53 immunopositivity (*). Coarse granular cytoplasmic phospho-p53 immunostaining (b) was replaced by a fine granular pattern in poorly perfused brain sections (e,f) illustrates the nuclei identified by Hoechst staining (blue). The inset is the merged image of (d,e and f). Scale bars: 10 µm.