Figure 1.

Mass Cytometry Analysis Reveals Four Phenotypically Basophilic Granulocyte Subpopulations Identified by Traditional CD45⁺HLA-DR⁻CD123⁺ Gating

(A) Gating strategy plots for total phenotypically basophilic granulocytes (PBGs, CD45⁺HLA-DR^-CD123⁺), eosinophils (CD45⁺HLA-DR⁻CD123⁻CD14⁻CD66b⁺CD193⁺Siglec8⁺), and neutrophils (CD45⁺HLA-DR^-CD123⁻CD14⁻CD66b⁺CD16⁺) using classic lineage markers.

(B) Implementation of Leiden clustering algorithm using the following markers: CD123, CD16, FcεRI, CD244, CD53, CD305, and CD193.

(C) Four donors' mean median expression (arcsinh with cofactor 5 transformation) across clusters identified by the Leiden algorithm, hierarchically clustered by protein expression and by Leiden clusters.

(D) Normalized single cell expression across of the following proteins: CD123, CD16, FcεRI, and CD244.

(E) CD16, FcεRI, and CD244 used to manually draw gates in biaxial plots to distinguish the four PBG subpopulations.

(F) The four-gated PBG subpopulations superimposed on the original tSNE plot coordinates. See also Figures S1 and S2.