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. Author manuscript; available in PMC: 2025 Apr 28.
Published in final edited form as: Nat Immunol. 2024 Jan 10;25(3):562–575. doi: 10.1038/s41590-023-01721-9

Fig. 3. Germinal centers disproportionately produce DP MBCs.

Fig. 3

a, Schematic for MBC generation with anti-CD40L blockade. b, Left: representative flow cytometry plots of NIP+ B cells (as gated in Supplementary Fig. 3a) produced in a, showing CD38hiFaslo MBC and CD38loFashi GCBC; Right: representative flow cytometry plots of MBCs gated from the Left panel. c, From left to right: number of GCs per spleen, percent of MBC subsets in total MBCs, and number of MBC subsets per spleen, based on flow cytometry gating in b. P-values were calculated using two-tailed Welch’s t-test. Bars display mean + s.d. Results are compiled from two-independent experiments (n=9 for each group). d, Mouse model for lineage tracing GC-derived memory cells. Mice were immunized with 100 ug NP-KLH in alum, and given 1 mg tamoxifen by oral gavage at days 9, 10, and 11 and euthanized at day 42. e, Representative spectral cytometry plots from splenocytes obtained from mice generated as in d, gated on NIP+ B cells (as in Supplementary Fig. 3c); arrows show cascading gating scheme.. f, from top to bottom: percent YFP+ cells among NIP+ GCBCs, percent of each MBC subset among YFP+ total MBCs, percent of each MBC subset among YFP− total MBCs, and percent YFP+ cells among NIP+ DP MBCs. Bars display mean + s.d. P-values were calculated using two-tailed paired t-tests. Results are compiled from two independent experiments (n=10 per group). g, Schematic of tamoxifen dosing window for 3 groups of mice given 1 mg of tamoxifen every other day from days 3–7, days 9–13, or days 3–13 and euthanized at day 42. h, top left: percent YFP+ cells among NIP+ GCBCs, top right: percent YFP+ cells among total NIP+ MBCs, bottom left: percentage of each MBC subset among NIP+ YFP+ total MBCs, bottom right: percent of YFP+ cells within each NIP+ MBC subset. Bars display mean + s.d. P-values were calculated using Mann-Whitney tests. Results are from one experiment for each time point (n=4 for days 3–7, n=4 for days 9–13, n=5 for days 3–13). *P <= 0.05, ***P < 0.001, ****P < 0.0001 for all plots. Actual p-values are listed in source data.