Figure 1. The Rroid locus controls peripheral group 1 ILC homeostasis.

A. Gene browser tracks of ATAC-seq (top) and RNA-seq (bottom) from indicated cell populations.

B. RNA fluorescence in-situ hybridization (FISH) of Rroid RNA in sort-purified CD19⁺ B cells and CD3, CD5⁻ NK1.1⁺ NKp46⁺ NK cells. Cells were probed for Rroid using Cy3-labeled probes (white). Nuclei were visualized with DAPI (blue).

C. Rroid expression in indicated mouse tissues was determined by quantitative PCR (qPCR). Normalized to Hprt expression. (n=3 mice per group). Data are representative of two independent experiments.

D. Rroid expression by qPCR in sorted cell populations from spleen, liver (Lv), small intestine lamina propria (SI), and lung parenchyma (n=3 mice per group; liver populations represent n=2 of 5 pooled mice each). Normalized to Hprt expression. Data are pooled from multiple independent experiments.

E. Representative flow cytometry plots of NK1.1⁺ NKp46⁺ cells in Rroid^+/+ and Rroid^−/− mice isolated from lung tissue. (Gated on live, CD45.2⁺ CD3, CD5⁻ cells).

F. Absolute numbers of CD3, CD5⁻ NK1.1⁺ NKp46⁺ cells in indicated mouse tissues (n=4 mice per group). Data are representative of three independent experiments.

G. Gating strategy (left) to identify CD49a⁺ ILC1s and CD49b⁺ NK cells in mouse liver. Cells were pre-gated on CD45.2⁺ CD3, CD5⁻ cells. Absolute numbers of liver and lung NK cells and ILC1s (right; n=4 mice per group). Data are representative of three independent experiments.

H. Absolute numbers of Lin⁻ CD45.2⁺ CD90.2⁺ Rorγt⁻ Eomes⁻ T-bet⁺ ILC1s in the small intestine intraepithelial lymphocyte (IEL) and lamina propria (SI-LPL) compartments (left) and salivary gland CD3, CD5⁻ CD45.2⁺ CD49b⁺ CD49a⁺ ILC1s (right). (n=3–4 mice per group). Data are representative of two to three independent experiments.

I. Absolute numbers of ILC2 and ILC3 populations from indicated tissues (n=3–5 mice per group). Intestinal ILC2s were Lin⁻ CD45.2⁺ CD90.2⁺ GATA3⁺ Rorγt⁻; ILC3s were Lin⁻ CD45.2⁺ CD90.2⁺ GATA3⁻ Rorγt⁺. Lung ILC2 were Lin⁻ CD45.2⁺ CD90.2⁺ T1/ST2⁺. Data are representative of three independent experiments.

*p≤0.05, **p≤0.01, ***p≤0.001. Two-tailed t-test. All error bars represent SEM. See also Figure S1.