Figure 5.

(A) Coimmunoprecipitation of VBC with Hs-CUL-2 but not with Hs-CUL-1 in vitro. Various combinations of the cDNAs encoding human F-VHL, elongin B, elongin C, Hs-CUL-1-HA, and HS-CUL-2-HA were expressed in a coupled transcription–translation system in the presence of [³⁵S]methionine. The translation products were immunoprecipitated with either M2 or HA antibodies and analyzed on a SDS/14% polyacrylamide gel followed by autoradiography. (B) Coimmunoprecipitation of Hs-CUL-2 with wild-type (wt) or mutant VHL (R167W, 157Δ) in vitro. cDNAs encoding human wt F-VHL, R167W, and 157Δ mutant VHL, elongin B, elongin C, and Hs-CUL-2-HA were expressed in a coupled transcription–translation system in the presence of [³⁵S]methionine. The translation products were immunoprecipitated with M2 antibody and analyzed on a SDS/14% polyacrylamide gel, followed by autoradiography.