Fig. 5

RNF31 regulates YAP protein stability via promoting YAP K48-linked poly-ubiquitination. A Western blot detecting of polyubiquitinated YAP was performed after coimmunoprecipitation in HEK293T cells treated in indicated. B-E Western blot detecting of K48-specific/K48R-specific/K63-specific/ mono-ubiquitinated YAP polyubiquitinated YAP was performed after coimmunoprecipitation in HEK293T cells treated in indicated. F-G Western blot detecting of polyubiquitinated YAP was performed after coimmunoprecipitation in HEK293T cells treated in indicated. H Schematic diagram to show lysine mutation site of YAP used in this study. I-J Ubiquitination of YAP at indicated multiple sites were measured by ubiquitination assay. K76 mutations (K76R) largely eliminated the ubiquitination effect of RNF31 on YAP protein. K Schematic illustration of RNF31 regulating Hippo/YAP signaling in TNBC progression. RNF31 protein associated with YAP and increased YAP protein degradation via inducing YAP K48-linked polyubiquitination, which repressed the Hippo/YAP axis activation and progression of TNBC cells. All the results are representative of three independent experiments