Fig 1: High-resolution fractionation of immature and mature basophils by scRNA-seq and FACS analysis.a UMAP projection of selected Basophils 2 hallmark genes. b Pseudotime analysis for selected clusters: Progenitors 1 (pink), Progenitors 2 (dark yellow), Basophil progenitors (dark green), Basophils 1 (brown), and Basophils 2 (cyan). Expression levels of selected Basophils 2 signature genes are shown. c Gene set variation analysis comparing Basophils 1 and Basophils 2. The top 5 differentially enriched pathways were shown. d FACS plots of bone marrow cells from Myb -68 GFP mice. FACS gating strategies are shown in Supplementary Fig.12. The top panel shows a plot of GFP+Lin- cells. Lin- indicates CD4-CD8-B220-Gr1-Ly6C-CD11c-cKit-Ter119-SiglecF-IL7Ra-NK1.1-. The left lower panel shows that GFP+Lin-FceR1a+ cells are subdivided into 4 groups: Basophil progenitors (CD34midLILRB4-FceR1ahigh, dark green), Basophils 1 (CD34lowLILRB4-FceR1ahigh, brown), early Basophils 2 (CD34lowLILRB4+FceR1ahigh, cyan), and late Basophils 2 (CD34-LILRB4+FceR1amid, blue). The right lower panel is a FACS contour plot showing LILRB4 and FceR1a expression levels of each subset defined in the left lower panel. In addition, data obtained from basophils in peripheral blood (red) were overlaid for comparison. Small black arrows inside plots show the direction of basophil differentiation. Representative plots from 3 independent experiments are shown. e May Grünwald Giemsa staining of each subset of basophil-lineage cells. Bar shows 20 µm.