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. 2017 Sep 4;7:10310. doi: 10.1038/s41598-017-10677-4

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© The Author(s) 2017

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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IL-10 expression. (A) ELISA revealed both AcES and TcES to be capable of increasing IL-10 secretion by canine PBMCs. For AcES this effect was more obvious at 15 µg/mL. In contrast, TcES-associated IL-10 secretion was stronger at 150 µg/mL and was only detected when cells were co-stimulated with LPS. (B) For analysis of intracellular IL-10 staining, PBMCs were gated on lymphocytes and monocytes based on side (SSC) and forward scatter (FSC) morphology. This revealed (C) lymphocytes but not (D) monocytes to be responsible for ES-induced IL-10 expression and the effect was independent from stimulation with LPS. P-values were calculated according to Wilcoxon signed rank test (ELISA without LPS stimulation) or paired t-test (ELISA with LPS stimulation, intracellular cytokine staining). Error bars show standard error of mean (SEM). Experiments were repeated six times at two different time points.