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Figure S1: Splenocytes prepared from naïve C57BL/6 mice or on Day 14 after heart transplantation were stained with Kd-PE, Kd-FTIC and Kb-APC, or stained with Kd-PE, Kd-FITC and streptavidin(SAV)-APC. All data are the mean and SEM of 3–4 mice, and no significant differences in the percentage of Kb-APC- or SAV-APC-reactive Kd Tet-DP B cells are observed between the naïve and transplanted mice (p > 0.05).

Figure S2: Background staining with sera from naïve C57BL/6 mice on BALB/c targets. Sera from naïve mice or 7 days after BALB/c heart transplantation were incubated with BALB/c splenocytes, and then stained with anti-mouse IgG-FITC and anti-mouse IgM-PE. Control samples had no sera added. Data are presented as mean fluorescence intensity ± SEM (three mice each group).

Figure S3: IRF4 and CD138 staining of plasmablasts identifies a B220lo IRF4hi CD138+ plasmablast population. IRF4lo are activated B cells comprising the prememory/GC as well as the memory and GC subsets. CD138+B220hi B cells are likely to be activated B cells but not committed PCs.

Figure S4: Detailed gating strategy for Figures 5 and 6. After gating on lineage negative DumpKd Tet-DP population, ≥95% of the cells are B220+, so further B220 gating is not necessary. The DumpKd Tet-DP cells are then subdivided into IgDhi and IgDlo populations and their expression of GC (GL7+Fas+) or other markers (not shown) were assessed.

ajt12350-sm-0002-SupTab-S1.docx12KTable S1: Germinal centers in lymph nodes from immunized and CTLA-4Ig treated mice.

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