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. 2018 Nov 13;8:16719. doi: 10.1038/s41598-018-34953-z

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© The Author(s) 2018

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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STAT3 phosphorylation analysis after stimulation. (a) Western blot analysis of whole cell lysates of PBMCs, unstimulated or 20 min. stimulated with 200 ng/ml IL6 or IL10. Expression of STAT3 phosphorylated at Y705 (pSTAT3) and total STAT3 (STAT3) of the two affected siblings and a healthy control was assessed; Actin as loading control. (b) Representative flow cytometric analysis showing diminished Y705-STAT3 phosphorylation after 20 min. stimulation with 200 ng/ml IL6 (solid line) versus unremarkable results after stimulation with 20 ng/ml IL10 (dotted line) and 10 ng/ml IL21 (dashed line) in lymphocytes of patient II.2 compared to unremarkable results in patient II.3 and a healthy control; filled gray area: unstimulated lymphocytes. (c) Flow cytometric analysis showing Y705-STAT3 phosphorylation after 20 min. stimulation with 20 ng/ml IL6 (solid line) or IL10 (dotted line) and 10 ng/ml IL21 (dashed line) comparable to healthy control in lymphocytes of one (representative of four) DOCK8-HIES patient. (d) Restored STAT3 phosphorylation after IL6 stimulation (solid line) in patient II.2 15 months after HSCT compared to unstimulated (filled gray area) and IL10-stimulated (dotted line) lymphocytes.