(A) Blimp1-GFP mice were immunized with NP-CGG/Alum, reimmunized with the same immunogen after 3 weeks and analyzed 4 days later. PCs were positively enriched from spleen and BM cells by CD138 PE magnetic bead-based selection and FACS sorted for qPCR analysis. Representative FACS plots of live Dump (CD4, CD8, F4/80, Ly6G)-B220+ cells with gates on TACI+CD98+ cells consisting of CD138+ Blimp1-GFPINT immature PCs or Blimp1-GFPHI mature PCs. (B)
Ltbr gene expression relative to Hprt among total spleen cells, CD8-Gr-1-F4/80-Dead- B220+IgD+ FO B cells, B220+PNA+GL7+IgD- GC B and PCs. FO and GC B cells were FACS sorted from spleens of WT mice immunized with NP-CGG/Alum for 11 days. (C) LTβR protein expression by flow cytometry in LTβR-deficient (Ltbrfl/fl Mb1-cre+/-) and WT (Ltbr+/+ Mb1-cre+/-) PCs. Data shown in all panels are representative of two independent experiments. Pooled data from two independent experiments are shown (n = 4–6 mice per group for A-B and n = 3 for C). * p < 0.05, ** p < 0.01.