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. 2021 Aug 10;12:4813. doi: 10.1038/s41467-021-25102-8

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© The Author(s) 2021

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 Representative FACS plots and quantifications of total GC B cells (gated on B220⁺ GL7⁺Fas⁺ splenocytes) from aged (24+wk) female C57BL/6 (WT) (black circles), female DKO(F) (red circles), male DKO(M) (purple squares), and male Yaa-DKO(M) mice (maroon squares). Data show mean ± SEM; n = 9 for WT, n = 15 for DKO(F) and Yaa-DKO(M), n = 14 for DKO(M) over 9 independent experiments; p-value by Brown–Forsythe and Welch ANOVA followed by Games–Howell’s test for multiple comparisons. b Representative immunofluorescence images of B220⁺ (blue) GL7⁺ (pink) GCs in spleens from the indicated mice. Data representative of at least 4 frames for at least 2 mice per genotype. Bars show 50 μm. c Representative FACS plots and quantifications of total PB/PC (B220^mid/loCD138⁺) in spleens of the indicated mice. Data show mean ± SEM; n = 9 for WT, n = 14 for DKO(F) and DKO(M), n = 15 for Yaa-DKO(M) over 9 independent experiments; p-value by Brown–Forsythe and Welch ANOVA followed by Games–Howell’s test for multiple comparisons. d Quantification of the FACS plots showing T_FH (CD4⁺ PD1^hiCXCR5⁺ Foxp3⁻) to T_FR (CD4⁺ PD1^hiCXCR5⁺ Foxp3⁺) ratio in spleens from the indicated mice. Data show mean ± SEM; n = 7 for WT, n = 12 for DKO(F) and DKO(M), n = 13 for Yaa-DKO(M) over 7 independent experiments; p-value by Brown–Forsythe and Welch ANOVA followed by Games–Howell’s test for multiple comparisons. e Quantification of FACS plots showing IFNγ⁺IL-21⁺ T cells in spleens of the indicated mice following 4 h treatment of splenocytes with PMA/Ionomycin. Data show mean ± SEM; n = 4 for WT and Yaa-DKO(M), n = 5 for DKO(F) and DKO(M) over 4 independent experiments; p-value by Brown–Forsythe and Welch ANOVA followed by Games–Howell’s test for multiple comparisons. f, g Representative FACS plots and quantifications of CD11c⁺ GL7⁺Fas⁺ cells (gated on B220⁺ splenocytes) (f) or CD11c⁺ PB/PC (B220^mid/loCD138⁺) (g) from the indicated mice. Data show mean ± SEM; n = 7 for WT, n = 12 for DKO(F) and Yaa-DKO(M), n = 3 for DKO(M) over 7 independent experiments; p-value by Brown–Forsythe and Welch ANOVA followed by Games–Howell’s test for multiple comparisons.