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. 2019 Nov 27;2:436. doi: 10.1038/s42003-019-0669-y

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© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 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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Fig. 2 — γ-Irradiation aggravates mammary tumorigenesis and promotes p53LOH in MMTV-ErbB2 mouse model. a, b Kaplan-Meier survival curves of irradiated and non-irradiated MMTV-ErbB2 mouse model. Single-dose of 5 Gy γ-irradiation at the time of onset of pre-malignant lesions (80 days) aggravates mammary tumorigenesis in p53H/+;ErbB2 vs. p53−/+;ErbB2 (p < 0.001 and p = 0.04, respectively) (a), but not in p53+/+;ErbB2 mice (p = 0.892) (b). n values are indicated in the figure and represent the number of mice. c Example of LOH analysis in tumors from p53H/+;ErbB2 mice. Non-irradiated mice are showing LOH in few mice only (top lanes 7–9). Irradiated mice, showing LOH in all but 1 mouse (bottom lane 17). d P53 expression in a panel of cell lines established from mammary tumors of MMTV-ErbB2 mice with different p53 genotypes. HSC70 is a loading control. e Mutp53 enhances LOH following γ-irradiation in cell culture (n = 3 independent samples Error bars represent ± SD). Cultivated mammary tumors cells were irradiated (9 Gy), or not, and grown up to 25 days post-irradiation. DNA was extracted at the indicated time points. The copy number of p53 wt and mut alleles was quantified by real-time PCR. DNA extracted from tail tissue samples of the corresponding genotype was used for copy number control. The experiment was repeated three times. Summary of a representative experiment. f Wtp53 retains transcriptional activity and, in response to Mdm2 inhibitor nutlin, induces its target p21 and Mdm2 in mutp53 heterozygous cells. Nutlin does not induce Mdm2 in p53H/H;ErbB2 and p53−/−;ErbB2 MECs. n = 3 independent experiments. Error bars represent ± SD.