Figure S5.

Analysis of recall responses in mice treated with anti-BAFF or following deletion of BAFFR. (A) WT mice were immunized with CGG or NP-CGG in alum on day 0, followed by anti-BAFF or isotype control antibody injections 46 d and 50 d later and rechallenged with NP-CGG 16 d later. Blood was withdrawn 45, 66, 73, 80, and 87 d after primary immunization, and spleen and bone marrow were analyzed at day 87. (B) Images of example ELISPOT wells showing NP⁺ IgM⁺ or IgG1⁺ antibody secreting cells in spleen and bone marrow from mice treated as indicated. Each spot corresponds to a single antibody secreting cell. (C) Mixed bone marrow chimeras were generated by reconstituting irradiated Rag2^−/−Il2rg^−/− mice with 10% bone marrow from Baffr^+/+RCE or Baffr^fl/flRCE B6 mice (H-2K^b+) and 90% bone marrow from WT BALB/c mice (H-2K^d+) for 41 d. Mice were immunized with NP-CGG in alum on day 0, given five daily tamoxifen injections starting on day 35, and rechallenged with NP-CGG in PBS on day 58. Spleen and bone marrow were taken for analysis on day 79. (D) Flow cytometric analysis showing gating of H-2K^b+ or H-2K^d+ splenic IgG1⁺ MBCs (CD138⁻B220⁺Fas⁻PD-L2⁺) and bone marrow IgG1⁺ MBCs (CD138⁻B220⁺PD-L2⁺) as well as splenic and bone marrow IgM⁺ and IgG1⁺ PCs (FSC^hiCD138^hiB220⁻). Red boxes indicate gates used to identify cell populations. FSC, forward scatter; SSC, side scatter.