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. 2022 Aug 2;5:781. doi: 10.1038/s42003-022-03743-9

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© The Author(s) 2022

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. 5 — a Example images of wildtype mouse astrocytes stained for SA-β-gal seven days after irradiation with 0 or 10 Gy, followed by counterstaining with nuclear fast red. Scale bar, 150 μm. b Quantification of the percentage of SA-β-gal positive cells. Results are pooled from 3 separate experiments, representing a total of 17 male and 23 female pups across 7 litters. Values were normalized to the Male 0 Gy condition of the same experiment, which was arbitrarily set at 1. Two-way ANOVA: dose p < 0.0001, sex p = 0.0455, interaction p = 0.3676. **p < 0.01, ****p < 0.0001 as indicated by bracket. Data are means +/− SEM (n = 17–23/sex/condition). c Number of Geminin positive nuclei per high-powered field. Shown are the means +/− SEM and each of the individual replicate points (n = 5/sex/condition). Two-way ANOVA: dose p < 0.0001, sex p = 0.3948, interaction p = 0.5561. *p = 0.0145 and **p = 0.0024 as indicated by bracket. d Correlation between Cdkn2a (p16) mRNA levels at 24 h after irradiation with 0 or 10 Gy and the percentage of SA-β-gal positive cells at 7 days after irradiation in male and female wildtype astrocytes (n = 10 male/8 female). e Correlation between Cdkn1a (p21) mRNA levels at 24 h after irradiation with 0 or 10 Gy and the percentage of SA-β-gal positive cells at 7 days after irradiation in male and female wildtype astrocytes. Male correlation: r = 0.28, p = 0.441, female correlation: r = 0.79, p = 0.0207 (n = 10 male/8 female). f Correlation between the Cdkn1a/Cdk2 mRNA ratio at 24 h after irradiation with 0 or 10 Gy and the percentage of SA-β-gal positive cells at 7 days after irradiation in male and female wild-type astrocytes. Male correlation: r = 0.39, p = 0.2608, Female correlation: r = 0.94 p = 0.0005 (n = 10 male/8 female). g Representative western blot images of p21 and Cdk2 in non-irradiated and irradiated male and female wild-type astrocytes at 24 h, and irradiated male and female wild-type astrocytes at 7 days. Molecular weight markers (MW): p21 (15 kDa), Cdk2 (30 kDa), α-Tubulin (50 kDa). h Correlation between p21 protein levels at 24 h after irradiation with 0 or 10 Gy and the percentage of SA-β-gal positive cells at 7 days after irradiation in male and female wild-type astrocytes. Male correlation: r = 0.33, p = 0.2909, female correlation: r = 0.67, p = 0.0003 (n = 12 male/24 female). i Correlation between the p21/Cdk2 protein ratio at 24 h after irradiation with 0 or 10 Gy and the percentage of SA-β-gal positive cells at 7 days after irradiation in male and female wildtype astrocytes. Male correlation: r = 0.20, p = 0.5418, Female correlation: r = 0.53, p = 0.0081 (n = 12 male/24 female).