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. 2016 Nov 21;36(14):2002–2013. doi: 10.1038/onc.2016.374

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Copyright © 2017 The Author(s)

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Persistent DNA double-strand breaks and apoptosis induced by irradiation. p53^+/− and p53^R172H/− mice were irradiated with 2 Gy and sacrificed at indicated time points. The testes of non-irradiated control and irradiated mice were harvested for analysis. (a and b) 53BP1 and γH2AX immunostaining of non-irradiated mice testes. 53BP1 pan-nuclear staining was observed in Asg, A1-A4 spermatogonia. White arrows: γH2AX condensed staining in leptotene and zygotene spermatocytes. Sex-bodies and spermatids expressed high γH2AX. (c and d) Time course of DSB induction indicated by 53BP1 and γH2AX immnunopositive foci signals. pIR: post irradiation. Yellow arrows: DSBs foci 6–8 h post irradiation. Purple arrows: DSBs foci 2–3 days post irradiation. White arrows: γH2AX foci staining in spermatids. (e) 53BP1/γH2AX co-labelling of testes from p53^R172H/− mice 2–3 days post 2 Gy irradiation. Purple arrows: co-expressing of 53BP1/γH2AX in DSBs foci. (a–e): Scale bars=20 μm. (f) Apoptosis induction indicated by cleaved caspase 3 IHC staining. Scale bar=50 μm. (g) Quantification of the number of apoptotic cells at various time points of p53^+/− and p53^R172H/− mice by counting cleaved caspase 3 positive cells. Slides were scanned by VSViewer system and 20 views of × 10 images per slide were captured for the counting. Data points represent the mean values for each group.