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. 2024 Oct 15;15:8890. doi: 10.1038/s41467-024-53222-4

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Pro B cells were FACS-sorted from mice of the indicated genotypes, and flow cytometry was used to determine the fraction of dead pro B cells (subG1) after sorting (d0) or in culture after 3, 5, and 7 days (d3, d5, d7) with IL-7. wild type wt (n = 5), Pidd1^-/- (n = 6), p21^-/- (n = 3), p53^-/- (n = 5), BCL2^tg (n = 4) and Mcl1^tg (n = 4). b Percentage of counted cells with more than 4 centrioles, determined by immunofluorescence with ɣ-Tubulin and CP110 antibody staining and Hoechst nuclear staining. wt (n = 6), Pidd1^-/- (n = 4), p21^-/- (n = 3), p53^-/- (n = 5), BCL2^tg (n = 3 d5, n = 4 d0, d3, d7) and Mcl1^tg (n = 4). c TO-PRO3 + /Annexin V + untreated or QVD-treated wt pro B cells were assessed via flow cytometry after 3,5 and 7 days of culture. n = 4; d Percentage of counted cells with more than 4 centrioles, determined by immunofluorescence with ɣ-Tubulin and CP110 antibody staining and Hoechst nuclear staining. n = 4; e Bar graph showing the percentage of wt (n = 5), p53^-/- and BCL2^tg (n = 3) pro B cells with 1,2 or 3 mature CEP164 + centrioles directly after sorting (d0) or after 3 days in culture (d3) with IL-7. CEP164 foci were determined by immunofluorescence with ɣ-Tubulin and CEP164 antibody and Hoechst nuclear stain. f Flow cytometry histograms of DNA marker TO-PRO3 of pro B cells expanded for 3 days with IL-7 and then treated with DMSO (UN) or 2, 4 µM DHCB for 48 h. g Fraction of cells in subG1. wt, Pidd1^-/-, BCL2^tg (n = 3). h Fraction of polyploid cells alive, wt, Pidd1^-/-, BCL2^tg (n = 3). Error bars in all panels represent the mean ± SD of biological replicates (individual mice). Genotypes were compared by two-way-ANOVA Tukey’s multiple comparisons test (g, h) and Bonferroni’s multiple comparisons test (a-b to compare to wt, and c, d). Source data and exact p-values (< 0.0001) are provided as a Source Data file.