Fig. 1: Gating strategy for isolation of ILC2s from lungs.
From: A diversity of novel type-2 innate lymphoid cell subpopulations revealed during tumour expansion
The gating of a standalone experiment to illustrate the gating strategy, which is the same for all ILC2 isolations in the study. ILC2 cells were enriched with EasySep Mouse ILC2 Enrichment Kit (STEMCELL Technologies) first then sorted from lungs of tumour bearing animals by FACS as Lin− ST2 + CD127 + Thy1.2+ cells. Sequential gating strategy is based on cell size [P1: small and non-granular cells, forward scatter (FSC) versus side scatter (SSC)], depletion of doublets (P2: SSC vs. Trigger Pulse Width), and selection of CD45 + cells. Cells with lineage-related markers were rigorously depleted during isolation process and the resulting cell numbers at each stage of purification is shown. The final gate (double positive for ST2 and Thy1.2) includes ILC2s isolated, enriched and sorted from lung tissue.
