FIG. 8.

The hydrophobic residues contained within the transactivation domain are required for the interaction with TBP. Immunoblot analysis was performed with the immunoprecipitation samples, untransfected control cells (lane 1) and cells transfected with 2 μg of pBK50b (lane 2) or pBK50M7 (lane 3). Polypeptides were resolved on an SDS–12% polyacrylamide gel and then transferred to nitrocellulose membranes. After transfer, the membranes were blocked, incubated with a 1/1,000 dilution of the anti-TFIID monoclonal antibody, washed, and incubated for 1 h at 37°C with a 1/1,000 dilution of anti-mouse immunoglobulin conjugated with horseradish peroxidase in blocking buffer. After five washes with PBS, the nitrocellulose membranes were developed, using enhanced chemiluminescence.