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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: Ajuba inhibits hepatocellular carcinoma cell growth via targeting of β-catenin and YAP signaling and is regulated by E3 ligase Hakai through neddylation

Fig. 6

Hakai associates with β-catenin and induces its translocation. a BEL7402 cells stably expressing Flag-tagged Hakai were fixed for immunofluorescence and stained for β-catenin protein (green) and DAPI (blue). Representative merged images showing overlap of fluorescence signals are shown. Scale bar = 25 μm. b Immunoblot analysis of β-catenin, Cyclin D1 and Hakai in BEL7402 cells stably expressing Hakai. GAPDH was used as a loading control. The ratios of expression Hakai and Cyclin D1 to their corresponding GAPDH are represented. Data are presented as Mean ± SEM from three independent experiments (***p < 0.001). c BEL7402 cells of stably expressing Hakai or vector control were transfected with two siRNA duplexex targeted to β-Catenin (siβ-Catenin) or control siRNA (siControl) for 48 h. Cell lysates were analyzed by immunoblotting for Hakai, β-catenin and Cyclin D1 protein expression. GAPDH was used as a loading control. d Co-immunoprecipitaion (IP) of GFP-tagged β-catenin and Myc-tagged Hakai or Myc-tagged Ajuba in 293 T cells. e IP of endogenous Hakai and Ajuba in BEL7402 cells with anti-Hakai antibody. f Co-IP of wild-type Hakai (WT) or its deletion mutants and endogenous β-catenin in transfected BEL7402 cells. IB, immunoblot. IP, immunoprecipitation. WCL, Whole-cell lysates. The results shown are representative of three separate experiments

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