(A) Experiment design. Itgax-DTR.BALB/c or Tg-negative littermates were used to make BM chimeras. cDCs were depleted with DT on days -1, -2, 1, 3 and 5. 9-10 × 106 AM14.BALB/c B cells were transferred into the mice on day 0, and activated with PL2-3 on days 0, 2 and 4. Spleens were harvested on day 6.
(B) Representative flow cytometry gating of live AM14 B cells by surface and intracellular 4-44 (anti-idiotype) staining, and further subgating for CD22lo, CD44hi plasmablasts.
(C) Number of AM14 B cells (surface and intracellular 4-44+) per spleen.
(D) Number of AM14 plasmablasts (4-44+, CD22lo, CD44hi) per spleen.
(E & F) ELISpot assays for the number of 4-44+, IgM+ or IgG2a+ AFCs per spleen. Data were combined from 2 experiments to obtain 4-8 mice per group. Bars represent mean and SEM. *p < 0.05; **p < 0.01 by ANOVA. See also Figures S1 and S2.