Figure 1. Phenotypic and morphological characterization of P. alecto BM-derived dendritic cells0 and macrophages.

(a) Histogram representation of the membrane expression of CD44, CD11b, CD172a (SIRPα), MHC-II (mouse anti-IA/IE antibody), CD3 and IgG by CD11b⁻CD3⁺ IgG⁺ T cells (red line), CD11b⁻CD3⁺IgG⁺B cells (violet line), and MHC-II⁻ (grey line) or MHC-II⁺ (black line) CD11b⁺ myeloid cells (see Supplementary Fig. S1a for the gating strategy). (b) Flow cytometry analysis of primary ex vivo bone marrow (BM, Day 0, upper panels) mononuclear cells or following 6 days (D6) in culture in the presence of FLT3L, GM-CSF + IL-4 or CSF-1. Non-doublet, live cells (see Supplementary Fig. S2 for the gating strategy) were first gated based on their size (FSC-A) and granulosity (SSC-A). CD44^hiCD11b⁺ cells were then analysed for CD172a (SIRPα) and MHC-II (anti-mouse I-A/I-E antibody) to define MHC-II⁻, MHC-II^int and MHC-II^hi cells. (c,d) Proportion of (c) CD11b⁺ cells among live cells or of (d) MHC-II^hi (upper panel), MHC-II^int (middle panel) and MHC-II⁻ (lower panel) cells, of ex vivo primary BM (D0, dashed filled histogram) or D6 cultured BM cells with FLT3L (white filled histogram), GM-CSF + IL-4 (grey filled histogram) or CSF-1 (black filled histogram). (e) Giemsa staining and (f) CD11b staining (red) observed by differential contrast/confocal microscopy, of D6 cultured BM cells with FLT3L (left panel), GM-CSF + IL-4 (middle panel) or CSF-1 (right panel). P values were calculated using the paired t-test.