Mediators of Inflammation / 2015 / Article / Fig 3

Research Article

CXCR6 Expression Is Important for Retention and Circulation of ILC Precursors

Figure 3

CXCR6 contributes to fetal ILC3 and ILC3 precursor circulation. (a) Flow cytometry of lineage depleted (Lin: CD3ε, CD11c, CD19, Ter119, Gr1, NK1.1) E15.5 fetal liver (FL). ILC3 precursors are defined as Lin  IL-7Rα+   RORγt+. (b) Percentages of Lin IL-7Rα+   (left panel) and absolute numbers (right panel) of FL ILC3 precursors as defined in Figure 2(a) in FL from Cxcr6+/+ (wt, blue losange), Cxcr6Gfp/+ (HZ, red square) or Cxcr6Gfp/Gfp (KO, green triangle) E15.5 embryos. (c) Flow cytometry of Lin compartment fetal spleen (FS) from Cxcr6Gfp/+ E15.5 embryos. Lin   IL-7Rα+ cells (red) and total Lin cells (filled gray) are analyzed for CXCR6-GFP (left histogram) and RORγt (right histogram). (d) Scheme depicting injection and reconstitution experiment. (e) Analysis of reconstitution experiment. 1000 to 2000 Ly5.2 Lin   IL-7Rα+ CXCR6+ cells were injected in nonlethally irradiated Rag2/γc/ Ly5.1 mice. Recipients were killed 4 weeks after injection and intestinal lamina propria (LP, upper panels) and liver (LV, lower panels) were analyzed by flow cytometry. (f) Percentages of reconstitution experiment as explained in Figures 2(d) and 2(e) using Cxcr6Gfp/+ (HZ, red squares) or Cxcr6Gfp/Gfp (KO) E15.5 embryos in intestinal LP (left panel) or LV (right panel). Results are representative of at least 3 experiments each ((a), (c): ), or are from 3 pooled experiments ((b), wt: , HZ: , KO: ), or are from at least 2 pooled experiments ((e), (f), LP HZ: , LP KO: , LV HZ: , LV KO: ). In ((b), (f)), each dot represents a single mouse. Statistical data are displayed with mean and SEM (Student’s unpaired bilateral test, ; ).