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. 2018 Aug 28;9:3486. doi: 10.1038/s41467-018-05852-8

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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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Fig. 4 — Wwox inhibits the interaction of JAK2 and STAT3. a Co-immunoprecipitation of Wwox and JAK2 shows Wwox–JAK2 interaction. 293T cells were transfected with plasmids expressing Myc-tagged full-length Wwox or HA-tagged full-length JAK2, as indicated. Whole-cell lysates were used in immunoprecipitation (IP) with anti-Myc antibody or anti-HA antibody. b Endogenous Wwox and JAK2 interact in vivo. Lysates of MCF-10A and T47D cells were subjected to IP with anti-JAK2 antibodies. c Wwox interacts with JAK2 mainly through the WW1 domain. IP and immunoblot (IB) of cell lysates from 293T cells expressing HA-tagged STAT3 and Myc-tagged Wwox or Myc-tagged truncated Wwox proteins (illustrated in Supplementary Fig. 5). Whole-cell lysates were immunoprecipitated with anti-HA and immunoblotted with anti-Myc antibody. d Wwox reduces p-STAT3 and p-JAK2 levels. MDA-MB-231 cells were transfected with Wwox plasmids and were treated with IL-6 for 30 min. The whole-cell lysates were immunoblotted with the indicated antibodies. e Interaction pattern of Wwox and JAK2 under IL-6 stimulation. IP and IB of cell lysates from 293T cells expressing HA-tagged JAK2 and Myc-tagged Wwox; cells were treated with IL-6 for the indicated time spans. f Wwox inhibits the interaction of JAK2 with STAT3. MCF-7 cells were transfected with Wwox siRNA, along with IL-6 stimulation; cell lysates were subjected to IP and IB analyses