Fig 1: Bone marrow-derived basophils (BMBAs) consist of two distinct subpopulations.Bone marrow cells were cultured in the presence of 0.3 ng/mL of IL-3 for 7 days. a CD200R3+cKit− BMBAs were gated (left panel) and the surface expression of FcεRIα and CD49b is shown (right panel). b, c Two subpopulations of BMBAs (FcεRIαhiCD49blo and FcεRIαloCD49bhi) were separately isolated and subjected to May-Grünwald-Giemsa staining (Scale bar, 10 μm) (b) and flow cytometric analysis (c). d–f BMBAs were subjected to scRNA-seq analysis. Shown are UMAP plot of basophil clusters (d), violin plots of the expression of indicated genes in each cluster (e), and volcano plot of genes differentially expressed in Baso1 versus Baso2/3 (f). Blue and pink dots indicate genes upregulated in Baso1 and Baso2/3 clusters, respectively. To calculate adjusted p value, non-parametric Wilcoxon rank sum test with Bonferonni correction was conducted. g CD200R3+cKit- BMBAs were subdivided into FcεRIαhiCD49blo and FcεRIαloCD49bhi subpopulations (left panel), and the surface expression of CLEC12A and CD9 in each subpopulation is shown (right panels). h CD200R3+cKit− BMBAs were subdivided into CLEC12AloCD9hi and CLEC12AhiCD9lo subpopulations (left panel), and the surface expression of FcεRIα and CD49b in each subpopulation is shown (right panels). Data shown in (a–c) and (g, h) are representative of at least three independent experiments. Data in (d–f) were obtained from a single experiment.