Fig. 1.

OX40L is highly expressed by intestinal ILC3s of adult mice. Splenocytes and small (SI) and large intestinal (LI) LPLs were isolated from 6-week-old (a–j and l) or young-age (k and l) Rag1^–/– mice. Cells were analyzed when freshly isolated (a–d) or after being cultured overnight (e-l). a, e, and k The expression of OX40L and RORγt gated on lineage-negative (Lin^–, CD3^–B220^–CD11b^–CD11c^–) cells in live lymphocytes was analyzed by flow cytometry. b and l Percentages of OX40L⁺ cells gated on ILC3s (Lin^–RORγt⁺) are shown. c The expression of OX40L in NCR⁺ILC3s (Lin^–RORγt⁺NKp46⁺), NCR^–ILC3s (Lin^–RORγt⁺NKp46^–CD4^–), and Lti cells (Lin^–RORγt⁺NKp46^–CD4⁺) was analyzed by flow cytometry. d Percentages of OX40L⁺ cells gated on indicated ILC3 subsets are shown. f Expression of Lin and OX40L gated on live lymphocytes was analyzed by flow cytometry. Histogram of RORγt expression gated on Lin^–OX40L⁺ cells is shown. g Percentages of Lin^–OX40L⁺ (upper left quadrant in (f)) and Lin⁺OX40L⁺ cells (upper right quadrant in (f)) gated on live lymphocytes are shown. h MFI of OX40L of Lin^–OX40L⁺ and Lin⁺OX40L⁺ cells in (f) is shown, respectively. Connected dots were paired data from biological repeats. Statistical analysis was performed with paired t test. i Percentages of RORγt⁺ cells in Lin^–OX40L⁺ cells in (f) are shown. j Percentages of the composition of different ILC3 subsets within OX40⁺ILC3s are shown. a–l Data are representative of three independent experiments. LPLs, lamina propria lymphocytes; MFI, mean fluorescence intensity