Fig. 2
From: Activation of DR3 signaling causes loss of ILC3s and exacerbates intestinal inflammation
Autocrine and paracrine GM-CSF induced by α-DR3 is critical for ILC3 loss. Rorcgfp/+ (a), Rag1–/–Rorcgfp/+ (b), or Rag1–/–Rorc-creRosa26stop-YFP mice (c) were treated with α-DR3, and large intestinal LPLs were analyzed 4 days later. a–c Expression of Ki67, Lin, RORγt, and YFP was analyzed. a, b Percentages of Ki67+ cells in ILC3s are shown. c Percentages of Lin–YFP+RORγt– and Lin–YFP+RORγt+ cells in Lin– cells are shown. d, e Rag1–/– mice were treated with α-DR3 antibody, and large intestinal LPLs were isolated for analysis 24 h later. f–h Mice were treated with α-DR3 with (α-DR3+α-GM-CSF) or without (α-DR3+IgG) injection of neutralization antibody for GM-CSF. i, j, l–n Rag2–/–Il2rg–/– mice were half-lethally irradiated and transferred with Thy1.1+ wild-type bone marrow (BM) and Thy1.2+Csf2–/– BM mixed at 1:1 ratio (i, j), Csf2–/– BM, or Csf2–/– BM and Rag1–/–Rorcgfp/gfp BM mixed at 1:1 ratio (l–n). i, l Protocols for bone marrow transfer. k Rag1–/– mice were treated with 10 µg of GM-CSF or control plasmid DNA through hydrodynamic injection, and large intestinal LPLs were isolated for analysis 4 days later. d–o Expression of Lin, RORγt, and GM-CSF in large intestinal LPLs was analyzed. d Percentages of GM-CSF gated on ILC3s (Lin–RORγt+) are shown. f, m Expression of RORγt gated on Lin– cells were analyzed. e, g, k, n Percentages of ILC3s (Lin–RORγt+) gated on Lin– cells are shown. j Percentages of ILC3s from wild-type origin (Lin–Thy1.1+RORγt+) and Csf2–/– origin (Lin–Thy1.1– RORγt+) in total lymphocytes are shown. h The total number of ILC3s are shown. The data are means ± SEM. o Small and large intestinal LPLs from 6-week-old littermate Rag2–/–Il2rg+/– and Rag2–/–Il2rg–/– mice were isolated, and expression of lineage markers and RORγt was analyzed. Percentages of ILC3s (Lin–RORγt+) gated on live lymphocytes are shown. c, n Statistical analyses were performed with paired t test. a–o The data are representative of at least two independent experiments. Source data are provided as a Source Data File
