Fig. 5.

Parkin regulates β-catenin by ubiquitination in ethanol-treated hepatic cells. a Confocal microscopy assay of primary hepatic cells from WT mice treated with ethanol or without ethanol. Cells were triple-stained with β-catenin antibody (Alexa 488; green), parkin antibody (Alexa 568; red), and DAPI (nuclear counterstain; blue) (Scale bars, 50 μm). b Interaction between parkin and β-catenin. HepG2 cells were transfected with Myc-vector or Myc-parkin. After ethanol treatment for 24 h, protein extracts were immunoprecipitated with anti-Myc agarose. c Ubiquitination of β-catenin following immunoprecipitation (IP) with an anti-HA antibody and immunoblot analysis for β-catenin, and IP with an anti-Flag antibody and immunoblot analysis for HA-ubiquitin. Flag-β-catenin-overexpressing HepG2 cells were transfected with HA-ubiquitin and/or parkin siRNA