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. 2018 Feb 5;8:2410. doi: 10.1038/s41598-018-20000-4

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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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MDM2 inhibitors induce a senescence-like state. (A,B) HCA2 fibroblasts were treated using the indicated concentrations of MI-63 (A) or nutlin-3a (B). p53 and p21 levels were analyzed by western blotting. Actin levels served as a loading control. (C) IMR-90 fibroblasts were transduced with a p53 luciferase reporter and treated with MI-63 or nutlin-3a. Extracts were prepared and analyzed by luminometry. (D,E) HCA2 cells were treated with MI-63 or induced to senesce by 10 Gy IR. SA-β-gal staining is shown in (D) and percentage of positive cells is shown in (E). (F,G) HCA2 cells were treated with nutlin-3a or induced to senesce by 10 Gy IR. SA-β-gal staining is shown in (F) and percentage of positive cells is shown in (G). (H-I) HCA2 cells were irradiated (IR) or treated with MI-63 or nutlin-3a, and immunostained for γH2AX. Representative images are shown in (H) and percentage of cells with >3 γH2AX nuclear foci is shown in (I). Data are representative of two independent experiments. ***p < 0.001 by 1-way ANOVA.