Figure 8.
Gating strategy for ILC3 characterization in C57BL/6 mouse. Lymphocytes were isolated from (A) spleen, (B) liver, and (C) small intestine. (D) Frequency and number of ILC3s (of CD45+ leukocytes) in spleen, liver, small intestine (SI), and lung. (E) SI lymphocytes were incubated with phorbol 12-myristate 13-acetate (PMA)/ionomycin and Brefeldin A for 3 hours. ILC3s (Thy1+Lin−TCR/CD3ϵ−CD127+Rorgt+) were analyzed for intracellular IL-17 and IL-22. (F) The frequency of IL-17- and IL-22-producing ILC3s. Data plotted are means ± SEM. Gating applied on live CD45+lin−Thy1+TCRs/CD3ϵ−CD127+RORγt+ cells. Lin cocktail included Ter119, CD19, B220, CD11c, CD11b, GR-1, and NK1.1. Numbers indicate percentages of cells in each gate.