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. 2021 Jan 19;89(2):e00533-20. doi: 10.1128/IAI.00533-20

FIG 3.

FIG 3

Identification of RORγt-enriched innate lymphoid cells (ILC3s) from Rag1−/− mouse intestinal lamina propria lymphoid cells. (a to c) The intestinal lamina propria lymphoid cell extract was labeled with vital dye and various antibodies as described in Materials and Methods and analyzed for live cells (a), lineage-negative cells (b), and CD45/CD90 double-positive cells (c). FSC, forward scatter. (d and e) Two adjacent cell clusters were identified from the CD45/CD90 double-positive cell population, and they were marked as non-ILC3s (representing 14.2% of the total lineage-negative lamina propria lymphoid cells) and ILC3s (30.1%), respectively, because the non-ILC3 cluster was 93.5% negative for the ILC3-specific transcriptional factor RORγt (d), while the ILC3 cluster was 97.2% positive for RORγt (e). Thus, RORγt-positive ILC3s can be purified from the CD45/CD90 double-positive cell population after excluding dead cells and lineage-positive cells using a flow cytometry cell sorter.