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. 2020 Jan 2;13(3):460–470. doi: 10.1038/s41385-019-0246-1

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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 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. 3 — a IL-10 expression in tonsillar lymphocytes 48 h following SAg-Sau stimulation. Representative dot plots with numbers in top left and right quadrants indicate the percentage of IL-10⁺ CD4⁻ and IL-10⁺ CD4⁺ T cells within total lymphocyte population. Results from 8 individual samples were analyzed and summarized in the bar chat. b Bar chart showing the percentage of IL-10⁺ cells within Foxp3⁺ CD4⁺ T cell and Foxp3⁻ CD4⁺ T cell populations, respectively, n = 3. c IL-10 concentration in the culture supernatant of 48 h-cultured MNCs either unstimulated (MC) or stimulated with SAg-Sau. Samples were measured by ELISA in duplicate, n = 7. d, e Tonsillar MNCs were stimulated with SAg-Sau CCS for 24 h following which CD4⁺ T cell isolation was performed. mRNA was extracted from isolated CD4⁺ T cells for RT-qPCR. The fold change in Il12a (d) and Ebi3 (e) mRNA expression compared to the media control (MC) are shown. Tests were performed in duplicate, n = 4. f, g Tonsillar MNCs were stimulated with SAg-Sau for 48 h and treated with brefeldin A for 4 h before harvesting cells for CD4⁺ T cell isolation. f Protein expression of IL-12A and EBI3 subunits in isolated CD4⁺ T cells. g CD4⁺ T cell lysate was immunoprecipitated with anti-IL-12A and blotted with anti-EBI3 to detect the expression of IL-35 heterodimer in CD4⁺ T cells. The blot image (f, g) are representative of 3 independent experiments. Data (a, b, c, d, e) were analyzed using paired t-test.