Figure 3.

Single antigen staining is sufficient to identify NadA-specific switched memory B cells. (A) Representative staining of NadA-specific memory B cells identified by single or dual antigen staining strategies with amine-labeled or sortagged NadA-Alexa fluorochrome conjugates. Gating of NadA-specific memory B cells was based on staining in naïve mice (overlay in light gray) and HSA specificity controls. (B) For each staining strategy, the percentage of NadA-specific memory B cells among total switched memory B cells is shown in immune (top) and naïve (bottom) mice after subtracting background HSA staining. Results are from three independent experiments performed with splenocytes pooled from four mice per group. Box plots depict median values and ranges; the box-crossing lines depict the means.