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. 2017 Sep 6;7:10739. doi: 10.1038/s41598-017-11208-x

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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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Growth suppressive effects of p53 or CD137L. (a) p53 ^+/+ calvarial osteoblasts or p53 ^−/− calvarial osteoblasts were cultured on plates. Cell proliferation was assessed at 48 h. Error bars represent SD (n = 3). *P < 0.05, Student’s t-test. (b) Immunohistochemical staining of Ki67 in mouse p53^+/+ or p53^−/− calvaria with or without radiation exposure. Representative images from three tissues in each group are shown. Scale Bars (left) = 50 μm, Scale Bars (right) = 20 μm. (c) A colony formation assay was performed. After ectopic expression of CD137L or mock, the number of SaOS2, U2OS or LM8 cells was determined. Whole cell extracts were subjected to western blotting with a human or mouse anti-CD137L antibody. Error bars represent SD (n = 3). **P < 0.001, Student’s t-test. β-actin (U2OS and SaOS2) and α-tubulin antibody (LM8) was used as loading control. (d) Cd137l (n = 3) or mock (n = 3) stable expression cell lines were inoculated into the left and right flanks of C3H mice; each group contains 2 mice. The tumour volume was calculated every 2 or 3 days. *P < 0.05, Wilcoxon rank-sum test.