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. 2020 Dec 17;10:22190. doi: 10.1038/s41598-020-79102-7

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© The Author(s) 2020

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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Effect of S. cerevisiae and Selemax on cytokine production by spleen and proportion of Tregs in inguinal lymph nodes. EAE mice received 14 oral doses of S. cerevisiae or Selemax. Total number of leukocytes from spleen adjusted per gram of tissue (a). Spleen cells (5 × 10⁶) were stimulated with MOG_35-55 (20 µg/mL) for 48 h and the production of IFN-γ (b), TNF-α (c), IL-2 (d), IL-17A (e), IL-6 (f) and IL-10 (g) were assessed in the supernatants. Representative gating strategy to assess regulatory T cells (Treg - CD25⁺Foxp3⁺) in inguinal lymph nodes (h). Total cells from inguinal lymph nodes were collected and analyzed by flow cytometry to determine the proportion of T helper cells (Th - CD3⁺CD4⁺) (i) and Treg in total Th cells (j). The proportion of T subsets was used to calculate the ratio between Treg and Th cells (k). Statistical analysis was performed by one-way ANOVA and subsequent Tukey’s test. All data were expressed as the mean ± SD and statistical differences were represented by *p < 0.05 and **p < 0.01. Two independent experiments were combined, n = 7–10 mice/group. The graphs were created using GraphPad Prism v.8.0.2, flow cytometry images were created using FlowJo 10.7.1 and image was modified with Adobe Photoshop v.22.0.0.