Mediators of Inflammation / 2015 / Article / Fig 2

Research Article

CXCR6 Expression Is Important for Retention and Circulation of ILC Precursors

Figure 2

CXCR6-deficient precursors are retained in the bone marrow. (a) Percentages in Lin (upper panels) and absolute numbers (lower panels) of BM CLP, ILCP, and ILC2P as defined in Figure 1(a) from Rag2/ Cxcr6+/+ (wt, blue losange), Cxcr6Gfp/+ (HZ, red square), or Cxcr6Gfp/Gfp (KO, green triangle) adult mice. (b) Histograms depicting levels of CXCR6-GFP in ILCP (left panel) or ILC2P (right panel) from Rag2/ Cxcr6+/+ (filled gray), Cxcr6Gfp/+ (red line), or Cxcr6Gfp/Gfp (green line) bone marrows. (c) Percentages of CXCR6+ cells (upper panels) and absolute numbers of (lower panels) BM ILCP (left panels) and ILC2P (right panels) in Rag2/ Cxcr6Gfp/+ (HZ, red square) or Cxcr6Gfp/Gfp (KO, green triangle) adult mice. (d) Cell culture of BM ILCP from Rag2/ Cxcr6Gfp/+ (upper panels) or Cxcr6Gfp/Gfp (lower panels) adult mice. Each population is sorted and cultured at 20 cells per well on OP9-DL4 stromal cells with IL-7, c-KitL, and Flt3-L for 7 days and analyzed by flow cytometry. Indicated obtained progenies were analyzed for T-bet, EOMES, RORγt, and GATA-3 expression. (e) Histograms of BM LSK cells (filled gray, selected as Lin IL-7Rα    Sca-1+) and CLP (red line) for Ki67 expression and DAPI levels. (f) Percentages of Ki67+ cells (upper panels) and DAPI+ cells (lower panels) among BM CLP, ILCP, and ILC2P in Rag2/ Cxcr6+/+ (wt, blue losange), Cxcr6Gfp/+ (HZ, red square), or Cxcr6Gfp/Gfp (KO, green triangle) adult mice. Data are representative of at least 3 experiments ((b), (e): ), or are from one experiment ((d), at least 4 wells of each conditions), or are from 3 pooled experiments ((a), (c), (f), wt: , HZ: , KO: ). In ((a), (c), (f)), each dot represents a single mouse. Statistical data are displayed with mean and SEM (Student’s unpaired bilateral test, ; ; ).