FIG. 2.

The binding of wild-type and mutant Rev proteins to hCRM1 in vitro. In vitro translated wild-type and mutant Rev proteins were immobilized on a resin and incubated with HeLa cell extracts in the absence (lane 2) or presence (lanes 3 to 5) of GTP-charged RanQ69L protein. The sample in lane 1 was incubated without cell extract and RanQ69L. After incubation, SDS-PAGE sample buffer was added to the supernatant (unbound fraction) and to the precipitated resin (bound fraction) for Western blot analysis. Chicken anti-hCRM1 antibody or mouse anti-GAL4 monoclonal antibody was utilized to detect hCRM1 or GAL4-fused proteins, respectively.