Fig. 3: Quantification of antigen-specific and total ASCs in BCMA-KOΔ3 and WT mice after secondary T-dependent immunization with NP-KLH.

A Schematic illustration of the experimental setup. BCMA-KOΔ3 and control mice were immunized with 100 µg NP-KLH in alum, boosted with 50 µg NP-KLH in PBS on day 42, and analyzed on day 128. B NP-specific IgG serum concentrations were determined by ELISA for wildtype (WT) (black, n = 7) and BCMA-KOΔ3 mice (gray, n = 8). C IgH-chain isotype-specific quantification of antigen (NP)-specific ASCs by ELISpot analysis in bone marrow. The images below are representative pictures of ELISpot analysis with numbers indicating the number of cells seeded per well (n = 6 WT and 11 KOΔ3 for NP-IgM; n = 7 WT and 11 KOΔ3 for NP-IgA; n = 7 WT and 8 KOΔ3 for NP-IgG). D Flow cytometric quantification of total ASC numbers per organ (bone marrow was pooled from one femur and one tibia/mouse, n = 7 WT and 11 KOΔ3). E IgH-chain isotype-specific quantification of total bone marrow ASCs by ELISpot analysis. The numbers below the ELISpot images indicate the number of seeded cells per well (n = 7 WT and 11 KOΔ3). F Stacked bar diagram with combined data from C and E. Bar diagrams show mean and SD with each dot indicating one mouse. Statistical analysis in B and F was performed using a two-way ANOVA with Šídák’s multiple comparisons test. Statistical analysis (C–E) was performed with unpaired t-tests, correcting for multiple comparisons by the false discovery rate (FDR) according to Benjamini, Krieger, and Yekutieli's Two-stage step-up Method. Exact p-values, mouse sex, and ages are provided in the Source Data file. ASC antibody-secreting cell, WT wildtype, ns not significant, i.p. Intra-peritoneal, mLN mesenteric lymph node, ^*p ≤ 0.05, ^**p ≤ 0.01.