CXCR6 Expression Is Important for Retention and Circulation of ILC Precursors
ILCP and ILC2P in the bone marrow heterogeneously express CXCR6. (a) Flow cytometry of lineage depleted (Lin: CD3ε, CD5, CD8, CD11c, CD19, TCRβ, TCRγδ, Ter119, Gr1, and NK1.1) adult bone marrow (BM) from Cxcr6Gfp/+ Id2Yfp/+ mice. Among Lin− IL-7Rα+ compartments are defined: CLP (filled gray, Flt3+ ), ILCP (red, Flt3− ), and ILC2P (blue, ). Each compartment is analyzed for CXCR6-GFP and ID2-YFP expression (bottom histograms). (b) Cell culture of BM ILCP, selected as ID2+ CXCR6− (upper panels) or ID2+ CXCR6+ (lower panels). Each population is sorted and cultured at 20 cells per well on OP9-DL4 stromal cells with IL-7, c-KitL, and Flt3L for 10 days and analyzed by flow cytometry. Indicated obtained progenies were analyzed for ID2-YFP, CXCR6-GFP, RORγt, and IL-7Rα expression. Results are representative of at least 3 experiments each ((a): ), or 2 experiments ((b), at least 4 wells of each condition).