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. 2017 Apr 21;7:1005. doi: 10.1038/s41598-017-00691-x

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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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SOCS2 regulates SOCS1, and SOCS1 depletion specifically promotes STAT1 but not STAT3 activation. (a) Effect of overexpression of SOCS2 on SOCS1 levels. HEK293 cells were transfected with constant amounts of SOCS1 plasmid and increasing amounts of SOCS2 plasmid. 48 hours post transfection, protein levels of SOCS1 and SOCS2 were measured by Western Blot. One representative experiment out of three is shown. (b) NOD1 and control silencing of iDCs was performed for 72 hours. Thereafter, cells were induced with IL-10 (30 ng/ml) for 15 minutes and pSTAT1, STAT1 and SOCS1 levels were analysed by Western Blot. One of two experiments is shown. (c,d,e) Immature DCs were transfected with siRNA directed against SOCS1 or a control siRNA. c Efficiency of SOCS1 silencing in iDCs was analysed by q-RT-PCR. (d) 72 hours post transfection, cells were induced with IL-10 (30 ng/ml) for 15 minutes and STAT1/3 phosphorylation was monitored using Western Blot analysis. To control for equal loading, total STAT protein was detected. One out of three independent experiments is shown. (e) Values of three independent experiments were used for quantification. Data represent mean and SD. For statistical analysis a one-way ANOVA with Tukey’s post hoc test was performed.