Figure 2

Interaction and co-localization of Hrd1 with CDV H protein. A, B Identification of the interaction between Hrd1 and CDV H protein by Co-IP. 293 T cells were co-transfected with Flag-CDV H and Myc-Hrd1 for 24 h, and Flag or Myc vector was used as the control. The transfected cells were lysed and immunoprecipitated with anti-Flag or anti-Myc antibody. The interactions were analyzed by Western blot with anti-Myc, anti-Flag and anti-GAPDH antibodies. C Identification of the interaction between Hrd1 and CDV H protein during viral infection by Co-IP. Vero cells were infected with CDV 851 (MOI = 0.1) for 72 h and then subjected to immunoprecipitation with anti-CDV H antibody. The interaction was analyzed by Western blot with anti-CDV H, anti-Hrd1 and anti-GAPDH antibodies. D Co-localization of Hrd1 with CDV H protein. 293 T cells were transfected with CDV H protein for 24 h. They were incubated with anti-Flag antibody and anti-Hrd1 antibody as primary antibodies and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG (green) and Alexa Fluor 594-conjugated AffiniPure Goat Anti-Rabbit IgG (red) as secondary antibodies. Nuclei were stained with DAPI (blue). Co-localization between CDV H and Hrd1 was confirmed by confocal microscopy. Bars: 10 μm.