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. 2024 Nov 4;15:9520. doi: 10.1038/s41467-024-51719-6

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Representative flow cytometry plots and (b) summary dot plots from BALB/c Rag2^−/− (n = 4) and BALB/c Rag2^−/− x Ctla4^−/− mice (n = 4) showing the proportion of CTLA-4-expressing cLP ILC1, ILC2, NKp46⁺ ILC3s and NKp46^- ILC3 cells. c Flow cytometry plot showing CTLA-4 in the lamina propria of the colon of BALB/c WT mice and C57BL/6 RORγt-eGFP mice. LPMC were stimulated with PMA, ionomycin and monensin for 4 h. d Representative flow cytometry plots and (e) summary dot plots from BALB/c WT mice (n = 4) and (f) BALB/c Rag2^−/− mice (n = 4) showing the proportion of CTLA-4-positive KLRG1 and NKp46 ILC upon stimulation with PMA and ionomycin. Both e and f used RM one-way ANOVA test with Holm-Sidak’s multiple comparison, where * P = 0.0215 and ** P = 0.0033 (g) KLRG1⁺ (n = 3) and (h) NKp46⁺ KLRG1^- cLP ILC (n = 3) were FACS sorted and cultured with OP9-DL1 for 48 hrs in the presence of rIL7, rIL2 and rIL-15 (unstim), with the addition of either PMA and ionomycin, rIL-12 and rIL-18, rIL-1β and rIL-23, rIL-25 and rIL-33, or rIL-12, rIL-18, rIL-1β, rIL-6, rIL-27 and rTGF-β (cocktail). The proportion of CTLA-4-positive KLRG1⁺ CD90.2⁺ and KLRG1^- NKp46⁺ CD90.2⁺ cLP ILC was measured upon seeding KLRG1⁺ and KLRG1^- cLP ILC, respectively. Both g and h used one-way ANOVA test with Holm-Sidak’s multiple comparison * P = 0.0111 ** P = 0.0015 **** P < 0.0001. All experimental n in Fig. 2 are biologically independent mouse samples.