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. 2018 Nov 9;8:16636. doi: 10.1038/s41598-018-35109-9

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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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ROCK2 but not ROCK1 interacts with pSTAT3 in both cytoplasmic and nuclear fractions of human T cells stimulated by TH17 skewing conditions. Human peripheral blood CD4⁺ T cells were treated with indicated doses of KD025 and then stimulated under TH17-skewing conditions for 2 hours. (a) Total cell extracts were prepared and analyzed by Western blot for pSTAT3 and pMLC expression. (b) Co-immunoprecipitation of pSTAT3 by anti-ROCK2 and anti-ROCK1 antibody in the nuclear fraction (c) Co-immunoprecipitation of pSTAT3 by anti-JAk2 antibody from total cell lysates of human T cells. (d) Co-immunoprecipitation of ROCK2 and ROCK1 with an anti-pSTAT3 antibody in both cytoplasmic and nuclear fractions. Co-immunoprecipitation of p300 (e) or JAK3 (f) by anti-ROCK2 and anti-ROCK1 antibody in the nuclear (e) or cytoplasmic (f) fractions. Numbers indicate ratio between the precipitated protein and the input (e). One representative of three different experiments is shown. Full-length gels are included in Supplementary Fig. 1.