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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. 6 — a Percentage of CD19+ or CD3+ lymphocytes in the peripheral blood (PBL) of Plk4^F/F or Mb1^Cre (n = 9, pooled controls), Mb1^Cre Plk4^F/F (n = 3) and Mb1^Cre Plk4^F/F Usp28^F/F mice (n = 4), determined by flow cytometry. b Fraction of pro/pre B, pro B, and large pre B cells in bone marrow. Control (Plk4 ^F/F or Plk4 Usp28^F/F) n = 5, Mb1^Cre Plk4^F/F n = 3, Mb1^Cre Plk4^F/F Usp28^F/F n = 3. c Fraction of splenic B cells of the genotypes listed in (b) was determined by flow cytometry. Serum IgM and IgG1 levels were determined by ELISA. d Serum anti-OVA-IgG1 and spleen weight of indicated genotypes were analyzed 13 days post-immunization. n = 4, as control homozygous and heterozygous Plk4 and Usp28^F/F mice were pooled. e Expansion microscopy (by factor 4) to determine centriole numbers in splenic B cells with acetylated Tubulin (AcTub) antibody and DAPI staining. Scale bars represent 2.5 µm. n = 96 cells per genotype. f Representative images (left) and number of CEP152 or ɣ-Tubulin foci (right) in CD19 + B cells of PBL by IF with CD19, CEP152, ɣ-Tubulin antibody and DAPI nuclear staining. n = 3. g Centriole depletion assessed by IF with ɣ-Tubulin, CP110 antibodies, and Hoechst staining in iGC B cells cultured on feeder cells with IL-4 for 4 days ± centrinone. n = 6 untreated (UN), n = 4 treatment. h DNA content analysis of iGC B cells (IL-4 for 4 days, + 4 days with either IL-4 or IL-21). n = 3. i Trim37 levels relative to Hprt were assessed by qRT-PCR in pro B and LPS-activated mature splenic B cells treated with 125 nM centrinone plus QVD. Data are shown as mean ± SD. n = 4. j qRT-PCR analysis of p53 target genes in untreated versus centrinone-treated pro B or LPS-treated splenic B cells. n = 4. Error bars in all panels represent mean ± SD (except error bars for spleen weight depict min to max) of biological replicates (individual mice). One-way-ANOVA Tukey’s multiple comparisons test (a–d, g) and Student’s two-tailed, unpaired t test (i, j except for Noxa two-tailed Mann-Whitney test was employed due to non-normal data distribution). Source data and exact p-values (< 0.0001) are provided as a Source Data file.