Fig 8. Gating strategy for identification of antigen-specific rabbit memory B cells.

Cells were analysed in a BD FACS ARIA III. A gate was drawn around the lymphocyte population (gated population represented 59.8% of events) (A). FSC-W and FSC-A were then used to eliminate doublets (gated population represented 95.7% of events) (B). 7AAD⁺ dead cells and T cells were eliminated in the “dump channel” (gated population represented 97.1% of events) (C). IgG⁺/ IgM^- B cells were identified and gated on (gated population represented 2.67% of events) (D). Finally, a gate (P1) was drawn around the double-positive mWISP-1 antigen-specific population (gated population represented 0.262% of events) (E). Cells from gate P1 were sorted into a 96-well PCR plate at either one or three cells per well for subsequent RT-PCR.