Fig. 3: DUB3 is a bona fide YAP1 deubiquitinase.
From: CK2α-mediated phosphorylation of DUB3 promotes YAP1 stability and oncogenic functions
A The direct interactions between bacterially purified GST-CK2α and His-YAP1 or His-PRMT6 were assessed using an in vitro interaction assay. Coomassie blue staining (CBS) was performed to quantify levels of GST and GST-CK2α. B A2780 cells were transfected with either vector (V), Flag-PRMT6 or Flag-YAP1. Anti-Flag affinity gel was used to immunoprecipitate Flag-YAP1 or Flag-PRMT6. Subsequently, a phospho-CK2 substrate antibody was used to assess the phosphorylation levels of YAP1 and PRMT6. C Flag-YAP1 was transfected into A2780 cells. Mass spectrometry analysis of YAP1-immunoprecipitates was used to identify potential YAP1 interactors. Cell lysates of A2780 and SKOV3 cells were immunoprecipitated using IgG, anti-DUB3 (D) or anti-YAP1 (E) antibodies. The endogenous interaction of YAP1-DUB3 was measured by immunoblotting. F The interaction of bacterially purified GST-YAP1and His-DUB3 was measured by an in vitro interaction assay. CBS was performed to examine GST and GST-YAP1 levels.
