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. 2018 Nov 1;9:4564. doi: 10.1038/s41467-018-07022-2

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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. 5 — TAK1 associates with and phosphorylates YAP/TAZ independent of LATS1/2. a Western blot analysis of YAP and TAZ in WT and LATS1/2 DKO HEK293A cells treated with TNFα and MG132 as indicated. b Western blot analysis of lysate of HEK293A cells transfected with Flag-tagged YAP and indicated kinases. c Phos-tag SDS-PAGE analysis of lysate of HEK293T cells transfected with Flag-tagged YAP and indicated kinases. d Western blot analysis of lysate of HEK293T cells transfected with Flag-tagged YAP or YAP5SA and indicated kinases. e Western blot analysis of lysate of WT and LATS1/2 DKO HEK293A cells transfected with Flag-tagged YAP-5SA mutant with indicated kinases. f Immunoprecipitation assay using anti-YAP antibody to detect endogenous complex of YAP and TAK1 in HEK293T cells. g Immunoprecipitation assay of Flag-tagged TAK1 or YAP protein expressed in HEK293T cells. h, i Wild-type, TAK1-KD and YAP purified from HEK293T cells (h) or recombinant GST-YAP from E.coli (i) were used for in vitro kinase assay. j Purified LATS1, wild-type and kinase-dead TAK1 from HEK293T cells, and recombinant GST-YAP were used for in vitro kinase assay. k Western blot analysis of cell lysate of wild-type and LATS1/2 DKO HEK293A cells transfected with wild-type or TAK1-KD and Flag-tagged YAP plasmids. All experiments were repeated three times independently