Figure 2.

Steps used to sort and verify identity of DEs, Bcon, and Tcon

(A) In this series of dot plots, consecutive gating was used to identify lymphocytes and exclude doublets and dead cells, respectively, among PBMCs stained with the four-antibody cocktail.

(B) Left dot plot (CD5 vs CD19) shows how gated live lymphocyte singlets are divided into CD5⁻CD19⁺ Bcon, CD5⁺CD19⁻ Tcon, and CD5⁺CD19⁺ subpopulations – The identities of CD5⁻CD19⁺ cells and CD5⁺CD19⁻ cells as Bcon and Tcon cells are confirmed by analysis each subpopulation for the lack or expression of the TCRαβ, respectively (not shown). Right dot plot (IgD vs TCRαβ) shows how gated CD5⁺CD19⁺ cells are used to identify DE subsets. DEs are divided into a major CD5⁺CD19⁺TCRαβ⁺IgD⁺ subset and a minor CD5⁺CD19⁺TCRαβ⁺IgD⁻ cells (contains class-switched DEs, see Ahmed et al. Ahmed et al. (2019) for details).

(C) This series of plots shows the steps used to verify identity of sorted DEs by re-running an aliquot of sorted DEs. Bcon, and Tcon cells. Numbers in quadrants indicate percentages.