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. 2023 May 22;24(7):1124–1137. doi: 10.1038/s41590-023-01519-9

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© The Author(s) 2023

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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Extended Data Fig. 1 — (a) Gating strategy for the analysis of GC B cells, centroblasts, centrocytes and Tfh cell populations in the iLNs of adult and aged BALB/c mice immunized with NP-KLH in alum. (b) Representative flow cytometry plots identifying GC B cells (Bcl6 + CD38⁻B220 + CD4-) in the inguinal lymph nodes (iLNs) of adult and aged BALB/c mice and their frequency (c) and total number (d) at the indicated timepoints post immunization. (e) Representative flow cytometry plots identifying Tfh cells (PD1 + CXCR5 + CD4 + Casp3-B220-) and their frequency (f) and total number (g) in the iLN at the indicated timepoints post immunization. (h) Representative flow cytometry plots identifying centroblasts (CXCR4^hiCD86^loBcl6⁺CD38⁻) and centrocytes (CD86^hiCXCR4^loBcl6⁺CD38^-) and their frequency and number (i-l) at the indicated timepoints after immunization. Adult and aged BALB/c serum antibody titers of NP20 (m) and NP7 (n) specific IgG1 and antibody affinity maturation (o) as determined by the ratio of NP7/NP20 IgG1 titers, at the indicated time point post immunization. The data is representative of two independent experiments (n = 14) where each symbol represents the mean ± SD and p-values were generated by performing a two-way ANOVA with Sidak’s multiple comparisons test.

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