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. 2021 Mar 2;14(4):923–936. doi: 10.1038/s41385-021-00386-7

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© The Author(s) 2021

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 — Mice were treated as described in Fig. 3a. a qRT-PCR analysis of Il17a mRNA expression in colon biopsies from non-infected and CR-infected WT mice treated or not with IL-33. Data from two independent experiments are shown as mean ± SEM of fold change induction over non-infected, PBS-treated group. b Flow cytometry dot plots of one representative infected colon showing the gating strategy applied to assess the proportion of IL-17A⁺ cells among Foxp3^-CD4⁺ T cells (right panel) within the colonic lamina propria. c Frequencies and absolute numbers of IL-17A⁺ cells were measured among Foxp3^-CD4⁺ T cells. d Percentage of ST2⁺ cells among Th17 cells. Bars represent the mean ± SEM of data from three independent experiments (n = 9–12 mice per group). All statistical analyses were performed using one-way ANOVA followed by Tukey’s multiple comparison test or Kruskal-Wallis ANOVA with Dunn’s multiple comparison test. e Sort-purified splenic CD4⁺CD25^- T cells were cultured for 5 days under Th17-polarizing conditions in presence or absence of IL-33. On day 5, cells were counted and stained for FACS analysis. The same gating strategy applied to colonic LPLs and shown in panel b was used in this particular dataset to measure the percentage and absolute numbers of IL-17A-expressing cells among Foxp3^-CD4 T cells (e), and to assess their mean fluorescent intensity (MFI) for IL-17A (f). In the same experiments, the percentage of Foxp3⁺ cells among CD4⁺ T cells was evaluated (g). Data from two independent experiments are shown (n = 6 mice). h Sort-purified CD4⁺CD25^- T cells were labeled with the proliferation dye eFluor 670 and cultured for 5 days under Th17-polarizing conditions with or without IL-33. Proliferation was determined as loss of eFluor 670 by flow cytometry. Representative histogram-overlay is illustrated. Bars indicate the mean ± SEM of n = 2 mice. Statistical analyses were performed using Paired t test. *P < 0.05; **P < 0.01; ***P < 0.001.