Figure 5.

Characterization of C57BL/6 mouse lung ILC2s with CD3ϵ/TCR separated from the Lin cocktail. (A) Lin-negative/low Thy1⁺ cells were gated, and TCR/CD3ϵ⁻ cells were further divided into Lin⁻ (green gate) and Lin^lo (bold black gate). ILC2s in each gate were identified by CD127⁺ST2⁺ and confirmed by GATA3⁺RORγt⁻ (right histograms). Lin⁺ population (red gate) was used as negative control. (B) Lung ILC2s were gated by Lin⁻/low Thy1⁺TCR/CD3ϵ⁻ST2⁺CD127⁺ and analyzed for CD25 expression. Lin⁺ population (red gate and red histogram) was used as negative control. (C) Lung lymphocytes were incubated with phorbol 12-myristate 13-acetate (PMA)/ionomycin and Brefeldin A for 3 hours. ILC2s (Thy1⁺Lin⁻TCR/CD3ϵ⁻ST2⁺CD127⁺) were analyzed for intracellular IL-5 and IL-13. Gating was applied on live, single-cell, CD45⁺ leukocytes. Numbers indicate percentages of cells in each gate.