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. 2020 Sep 7;13:451. doi: 10.1186/s13071-020-04332-7

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© The Author(s) 2020

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — Analysis of CD4⁺ T cell subsets in mice after fucoidan treatment. a Immunofluorescence of CD4 (green) and Foxp3 (red) in the liver granulomas from the normal or S. japonicum-infected mice treated with or without fucoidan. Original magnification: ×400. b The mean optical density of CD4⁺Foxp3⁺ cells in each group was digitized and analyzed using Image-Pro Plus software. c Gating strategy for CD3⁺CD4⁺Foxp3⁺ (Treg), CD3⁺CD4⁺IL-17⁺ (Th17), CD3⁺CD4⁺IFN-γ⁺ (Th1) and CD3⁺CD4⁺IL-4⁺ (Th2) cells. Percentages and absolute numbers of Treg (d), Th17 (e), Th1(f) and Th2 (g) cells from hepatic or splenic lymphocytes of each group were analyzed by flow cytometry. Data are expressed as the mean ± SD for each group (n = 5 for each group), and all experiments were performed twice with similar results. *P < 0.05, **P < 0.01, ***P < 0.001. Abbreviation: ns, not significant (ANOVA/LSD). Scale-bars: a, 25 µm