Figure 1. Atg5 is essential for innate lymphocyte development.

(A) Representative flow cytometric plots of lineage marker negative (Lin⁻) NKp46⁺NK1.1⁺ cells and (B) Lin⁻NK1.1⁻CD127⁺CD90.2⁺Rorγt⁺ (ILC3s) in indicated peripheral organs of NKp46^iCre × Atg5^fl/fl mice (“NK-Atg5^−/−”) or littermate Atg5^fl/fl controls (called “WT”) (C) Absolute numbers of mature NK cells (mNK; Lin⁻NK1.1⁺NKp46⁺DX5⁺Eomes⁺), ILC1 (Lin⁻NK1.1⁺NKp46⁺DX5⁻Eomes⁻), and (D) ILC3s in indicated peripheral organs of WT or littermate NK-Atg5^−/− mice (E,F) WT mice (CD45.1 × CD45.2) were lethally irradiated and reconstituted with an equal number of bone marrow cells from WT (CD45.1) and Ubc^Cre-ERT2 × Atg5^fl/fl (CD45.2) mice. (E) Schematic of experiment. (F) Both WT (CD45.1) and Ubc^Cre-ERT2 × Atg5^fl/fl (CD45.2) CHILP (Lin⁻NK1.1⁻DX5⁻FLT3⁻CD127⁺α₄β₇⁺CD25⁻) and ILC2P (Lin⁻NK1.1⁻DX5⁻FLT3⁻CD127⁺α₄β₇⁺CD25⁺) cells were adoptively transferred into sub-lethally irradiated Rag2^−/− × Il2rg ^−/− hosts and injected i.p. with 1mg of tamoxifen daily for 5 days. After 28 days, the SI and adipose tissue were harvested from recipient mice and analyzed for mature ILC2 (Lin⁻NK1.1⁻DX5⁻CD127⁺CD90.2⁺GATA3⁺KLRG1⁺) and ILC3 cells. (Lin⁻ refers to TCRβ⁻CD19⁻CD3ε⁻Ly6G⁻TER119⁻TCRγδ⁻ cells). Data are representative of two independent experiments, with n=5 per time point. Samples were compared using an unpaired, two-tailed Student’s t test, and data presented as the mean ± s.e.m. (***p < 0.001). See also Figure S1.