Mass cytometry antibody panels, normalization, and gating strategy. (A) Mass cytometry panel used for analysis in Fig. 1 A, Fig. 2 B, and Fig. 7 C. (B) Pre- and postnormalization plots of mass cytometry data used for Fig. 1, which is representative of the mass cytometry data used in Figs. 2 and 7. (C) Gating strategy of mass cytometry data used to identify live CD19⁺ B cells. Cleanup residual, center, offset, and length gates were not used for the data displayed in Fig. 2. (D) Flow cytometry plots demonstrating identification of T1 and T2 cells as CD27⁻IgD⁺CD10⁺ cells that are CD24⁺⁺⁺/CD38⁺⁺⁺ and CD24⁺⁺/CD38⁺⁺ respectively, T3 cells as CD27⁻IgD⁺CD10⁻R123^hi, and naive (N) B cells as CD27⁻IgD⁺CD10⁻R123^lo. CSM, class-switched memory. (E) SPADE trees demonstrating expression of IgM and CD45RB in the concatenated GALT sample (see also Fig. 2 A). (F) Heatmap demonstrating the median expression of panel markers from metaclusters 3 and 4 displayed in Fig. 2 B. SSC-A, side scatter area; SSC-W, side scatter width.