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. 2014 Mar 3;105(4):473–480. doi: 10.1111/cas.12359

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© 2014 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.

This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Fig. 3 — Effects of 2-methoxystypandrone (2-MS) on Janus kinase 2 (JAK2) and activities of other human kinases. (a) HeLa cells were incubated with indicated concentrations of 2-MS for 2 h before stimulation with interleukin-6 (IL-6) (30 ng/mL) for 15 min. Whole cell lysates were processed for western blot analysis and probed with anti-P-JAK2 (Y1007/1008), anti-JAK2 or anti-GP130 antibodies. Anti-α-Tubulin antibody was used as a loading control. (b) HeLa cells were incubated with indicated concentrations of 2-MS for 2 h before being processed for western blot analysis. (c) In vitro kinase assay of JAK2. JAK2 protein immunoprecipitated from HeLa cells was subjected to in vitro kinase assay in the presence of indicated concentrations of 2-MS. (d) Effects of 2-MS on activities of human kinases. In vitro kinase assays were processed by Millipore Kinase Services. (e) In vitro kinase assay of IKKβ. IKKβ protein immunoprecipitated from HEK293 cells was subjected to in vitro kinase assay in the presence of indicated concentrations of 2-MS.