Fig. 3

IL-23 is important for α-DR3-induced loss of ILC3s. a The mRNA expression of Csf2ra and Csf2rb in purified CD11b⁺ cells and ILC3s (Lin^–GFP⁺) from large intestinal LPLs of Rag1^–/–Rorc^gfp/+ mice was analyzed by real-time RT-PCR. b The cDNA product from real-time RT-PCR analysis in (a) was analyzed by electrophoresis. c Rag1^–/– mice were treated with α-DR3 antibody. The mRNA expression of Il23a and Il12b in large intestinal LPLs was analyzed by real-time RT-PCR. d–j Rag1^–/– mice were treated with α-DR3 in the presence or absence (control IgG) of neutralizing antibody for p40, p19, or p75. Large intestinal LPLs were isolated for analysis on day 4 post α-DR3 treatment. k, l Rag1^–/– mice were treated with 10 µg of IL-12, IL-23, or control plasmid DNA through hydrodynamic injection, and large intestinal LPLs were isolated for analysis 4 days later. m Rag1^–/– mice were treated with 10 µg of GM-CSF or control plasmid DNA through hydrodynamic injection with (GM-CSF+α-p40) or without (GM-CSF+IgG) i.p. injection of neutralization antibody for p40 simultaneously. n, o Littermate mice with indicated genotypes were treated with 10 µg of IL-23 or control plasmid DNA through hydrodynamic injection. k–o Large intestinal LPLs were isolated for analysis 4 days later. d–o Expression of Lin and RORγt were analyzed by flow cytometry. d Expression of RORγt gated on Lin^– cells is shown. e, g, h, k, m, n Percentages of ILC3s (Lin^–RORγt⁺) gated on Lin^– cells are shown. f, i, j, l, o The total numbers of ILC3s are shown. a, c, e–h, k–o The data are means ± SEM. i, j Dots indicate average number of ILC3s per mouse from every batch of the experiment. The data from the same batch of the experiment are connected with a solid line. 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