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. 2022 Sep 29;13:5730. doi: 10.1038/s41467-022-33326-5

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a–d Irf1^+/– and Irf1^–/– mice were challenged with an IL-23-expression vector for three days. a The expression of Ifng, Il22 and Il17a in gut tissue samples was quantified by qRT-PCR with total RNA (Il17: Irf1^+/– untreated n = 6, IL-23 n = 11; Irf1^–/– untreated n = 5, IL-23 n = 11; Ifng: Irf1^+/– untreated n = 6, IL-23 n = 11; Irf1^–/– untreated n = 6, IL-23 n = 11; Il22: Irf1^+/– untreated n = 5, IL-23 n = 10; Irf1^–/– untreated n = 5, IL-23 n = 11) and (b) serum-concentrations of IFN-γ, IL-22 and IL-17a were measured by specific ELISAs (IL-17: Irf1^+/– untreated n = 7, IL-23 n = 11; Irf1^–/– untreated n = 6, IL-23 n = 10; IFN-γ: Irf1^+/– untreated n = 7, IL-23 n = 11; Irf1^–/– untreated n = 6, IL-23 n = 10; IL-22: Irf1^+/– untreated n = 6, IL-23 n = 11; Irf1^–/– untreated n = 6, IL-23 n = 10). c, d Intestinal LPMCs were isolated and analyzed by flow cytometry. Graphs show frequencies of Tbet⁺ ILCs (lin^-Thy1.2⁺Tbet⁺ cells), ILC1s (lin^-Thy1.2⁺Tbet⁺EOMES^-NK1.1⁺Nkp46⁺), ROR-γt⁺ ILC3s (lin^-Thy1.2⁺RORγt⁺ cells) and of ILC3s expressing IL-22 (Irf1^+/–: n = 4, Irf1^–/–: n = 3). e–h Irf1^+/– and Irf1^–/– ILCs were sort-purified from LPMC before in vitro stimulation (n = 3/group). e Numbers of input cells and the yield of sort-purified ILCs (CD45⁺B220^-CD3^-CD5^-CD11b^-CD11c^-KLRG1^-CD127⁺Thy1.2⁺ cells) are shown. f–h ILCs were stimulated with IL-1β (20 ng/ml) and IL-23 (20 ng/ml) or left unstimulated (medium). f Total RNA of ILCs was analyzed by specific qRT-PCR. g After 72 h, supernatants were collected to measure concentrations of IL-17A, IL-22 and IFN-γ by ELISA. h IL-22 production of RORγt⁺ILCs was studied by flow cytometry. Data is expressed as mean ± SEM. Exact p-values defined by 2way ANOVA with Tukey´s multiple comparisons test are provided in the plots. n.d.: not detectable. Source data are provided as a Source data file.