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. 2023 Sep 22;14:1198310. doi: 10.3389/fimmu.2023.1198310

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Copyright © 2023 Sadeghalvad, Khijakadze, Orangi and Takei

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Gating strategy for ILC3 characterization in C57BL/6 mouse. Lymphocytes were isolated from (A) spleen, (B) liver, and (C) small intestine. (D) Frequency and number of ILC3s (of CD45⁺ leukocytes) in spleen, liver, small intestine (SI), and lung. (E) SI lymphocytes were incubated with phorbol 12-myristate 13-acetate (PMA)/ionomycin and Brefeldin A for 3 hours. ILC3s (Thy1⁺Lin⁻TCR/CD3ϵ⁻CD127⁺Rorgt⁺) were analyzed for intracellular IL-17 and IL-22. (F) The frequency of IL-17- and IL-22-producing ILC3s. Data plotted are means ± SEM. Gating applied on live CD45⁺lin⁻Thy1⁺TCRs/CD3ϵ⁻CD127⁺RORγt⁺ cells. Lin cocktail included Ter119, CD19, B220, CD11c, CD11b, GR-1, and NK1.1. Numbers indicate percentages of cells in each gate.