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. 2017 Jun 21;7:3974. doi: 10.1038/s41598-017-04322-3

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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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DC generation from monocytes with anti-inflammatory cytokines exhibited tolerogenic phenotype and produced anti-inflammatory cytokines. TolDC and C5-DC were harvested after maturation and characterized by flow cytometry, anti-CD14, anti-CD83, anti-CD80, anti-CD86 anti-B7-H1 and anti-B7-DC antibodies were used. (a) Histogram, (b) dot chart of MFI ratio tolDC to C5-DC. Data represent 26 independent experiments from 9 donors (CD86: n = 8). Wilcoxon signed-rank test was used to determine p values. (c) IL-10, IL-12p70, TGF-β1 and IFN-γ quantification in DC supernatants using BD CBA Flex Set and ELISA. These data represent 10–19 independent experiments from 9 donors. Culture medium plus maturation cocktail without cells were used as control. Turkey test was used to determine the statistical significance. ***p < 0.001. RNA was isolated from tolDC and C5-DC, and quantified by RT-PCR, (d) Relative RNA level of IL-35 and related genes, IL-10, TGFB1 in tolDC, C5-DC as control. ACTB, G6PDH or CYPB mRNA were determined from the same sample and used as endogenous reference genes. These data represent 7 independent experiments from 6 donors. Wilcoxon signed-rank test was used to determine p values. Error bars: s.e.m.