ARF alters the composition of the VHL30 complex.
(A) Protein components of the FLAG-VHL30 complex in the presence or absence of ARF co-expression.
293T or U2OS cells were transfected with FLAG-VHL30 alone or together with ARF. Forty-eight hours after transfection, the FLAG-VHL30 complex was purified by anti-FLAG immunoprecipitation and was analyzed by SDS-PAGE and silver staining. Notable differences are indicated by the arrows.
(B) ARF disrupts the binding of VHL30 and elongin C or cullin 2.
U2OS cells were transfected with FLAG-VHL30, FLAG-VHL19, and ARF, as indicated. Forty-eight hours after transfection, the interaction of FLAG-VHL with elongin C or cullin 2 was examined by anti-FLAG immunoprecipitation followed by anti-elongin C or anti-cullin 2 immunoblotting.