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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. 7 — a Example images of Nf1−/− DNp53 Cas9 control and p21 knockdown (KD) male and female astrocytes stained for SA-β-gal 5 days after irradiation with 0 or 8 Gy, followed by counterstaining with nuclear fast red. Scale bar, 150 μm. b–d Quantification of the percentage of SA-β-gal positive cells 5 days post-irradiation with 0, 3, 6, or 8 Gy. Values were normalized to the corresponding Male 0 Gy condition of the same cell line, which was arbitrarily set at 1. b Male vs female Cas9 control cells—two-way ANOVA: dose p < 0.0001, sex p < 0.0001, interaction p = 0.2093. c Male Cas9 vs male p21 KD - two-way ANOVA: dose p < 0.0001, genotype p = 0.1459, interaction p = 0.8529; d Female Cas9 vs female p21 KD: two-way ANOVA: dose p < 0.0001, genotype p = 0.0093, interaction p = 0.5635. Male Cas9 curve (blue) is shown for comparison. Open symbols are the means of the individual data points, which are presented as closed symbols (b–d n = 6–9/sex/dose). e–g Quantification of the percent of Nf1−/− DNp53 Cas9 control versus p21 knockdown (KD) cells with >10 γH2AX foci at 1, 6, 24, and 48 h following irradiation with 3 Gy (top row) or 8 Gy (bottom row). Two-way ANOVAs: e Male Cas9 versus female Cas9 at 3 Gy: time: p < 0.0001, sex: p = 0.079, interaction: p = 0.625; male Cas9 versus female Cas9 at 8 Gy: time: p < 0.0001, sex: p = 0.0014, interaction: p = 0.0524. f Male Cas9 versus p21 KD at 3 Gy: time p < 0.0001, genotype: p = 0.356, interaction: p = 0.775; male Cas9 versus p21 KD at 8 Gy: time p < 0.0001, genotype: p = 0.0645, interaction: p = 5773 g female Cas9 versus p21 KD at 3 Gy: time p < 0.0001, genotype: p = 0.205, interaction: p = 0.774; female Cas9 versus p21 KD at 8 Gy: time p < 0.0001, genotype: p = 0.006, interaction: p = 0.0418. Open symbols are the means of the individual data points, which are presented as closed symbols (e–g n = 3/sex/timepoint).