Figure 3.

hMSH4 interacts with the VBP1-VHL-p97 complex. A) Interaction between hMSH4 and VBP1 was analyzed in HEK293T cells by reciprocal coimmunoprecipitation (co-IP) as indicated. B) VHL and p97 coexist in the hMSH4 immunocomplex. HEK293T cells transfected with empty vector or Myc-hMSH4 were subjected to immunoprecipitation using an anti-Myc antibody. C) VBP1 modulates the interaction between hMSH4 and p97. HEK293T cells were first transfected with control shRNA or VBP1 shRNA sh-2 to knock down VBP1 and were then transfected with Myc-hMSH4. At 24 h post-transfection, cells were treated with MG-132 and subjected to immunoprecipitation using an anti-Myc antibody, and immunoblotting was carried out as indicated. D) HEK293T cells were transfected with p97 siRNA using Lipofectamine 2000 before transfection with Myc-hMSH4. The protein level of hMSH4 was examined by immunoblotting.