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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. 2 — a Irradiation dose response curves in male and female Nf1−/− DNp53 astrocytes. Curves represent combined values from five separate established cell lines—each consisting of a corresponding male and female line generated at the same time. Male IC₅₀ = 5.567 (95% CI 5.094–6.088), female IC₅₀ = 4.694 (95% CI 4.349–5.061). Two-way ANOVA: dose p < 0.0001, sex p = 0.0123, interaction p = 0.6034. Open symbols are the means of the individual data points, which are presented as closed symbols. Lines are non-linear fits to the individual data points (n = 5/sex). b Male and female Nf1−/− DNp53 cell growth curves after irradiation with 0, 3, or 9 Gy. Cell growth was tracked using live cell imaging; images were taken every 4 h for 3 days and used to calculate percent confluence over time. Two-way repeated measures ANOVA—male 0 Gy vs male 3 Gy: time p < 0.0001, dose p = 0.3394, interaction p = 0.9087; male 0 Gy vs male 9 Gy: time p < 0.0001, dose p = 0.0325, interaction p < 0.0001; female 0 Gy vs female 3 Gy: time p < 0.0001, dose p = 0.0183, interaction p < 0.0001; female 0 Gy vs female 9 Gy: time p < 0.0001, dose p = 0.05, interaction p < 0.0001. Open symbols are the means of the individual data points, which are presented as closed symbols. Lines are non-linear fits to the individual data points (n = 3/sex). c Normalized colony counts from the clonogenic assay in male and female Nf1−/− DNp53 cells 5 days after irradiation with 0, 4, 8, 12, or 16 Gy. Two-way ANOVA: dose p < 0.0001, sex p = 0.002, interaction p = 0.111. Open symbols are the means of the individual data points, which are presented as closed symbols. (n = 3/sex). d Representative immunofluorescence images of male and female Nf1−/− DNp53 astrocytes stained for γH2AX 0, 1, 6, 24, and 48 h after irradiation with 3 Gy. Scale bar, 20 μm. e Quantification of the percent of cells with >10 γH2AX foci at each timepoint in male and female Nf1−/− DNp53 astrocytes following irradiation with 3 or 8 Gy. Two-way ANOVA—male 3 Gy vs female 3 Gy: time p < 0.0001, sex p = 0.0004, interaction p = 0.0251; male 8 Gy vs female 8 Gy: time p < 0.0001, sex p = 0.0133, interaction p = 0.2784; female 3 Gy vs female 8 Gy: time p < 0.0001, dose p = 0.0168, interaction p = 0.2959.