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. 2020 Feb 21;16(8):1335–1348. doi: 10.7150/ijbs.41130

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The direct interaction between TBC1D25 and TAK1. (A) The co-localization of TBC1D25 and TAK1 in 293T cells. The staining of TBC1D25 is green, TAK1 is red and DAPI is blue in representative confocal images. (B-C) The TBC1D25 and TAK1 interaction was detected by Immunoprecipitation (IP) assays. 293T cells were co-transfected with Flag-TBC1D25 and HA-TAK1. 48 hours later, harvested cell lysates were subjected to IP with antibodies against Flag (B) or HA (C). (D-E) The direct interaction between TBC1D25 and TAK1 were verified by GST pull-down assays. GST-tagged proteins were incubated with immunopurified Flag-tagged proteins and then immunoblotted. (F-G) Co-IP of the full-length and truncated mutants of TBC1D25 (F) or TAK1 (G) using a HA or Flag antibody, respectively.