Figure 1.

Interaction of TSG101 with p53 and MDM2. Constructs expressing human TSG101, p53, or MDM2 proteins (2 μg of DNA for each plasmid) were introduced by transfection into Saos-2 cells, as indicated in Materials and Methods. Protein extracts from transfected cell populations were immunoprecipitated by the antibodies indicated. IP, immunoprecipitation; IB, immunoblotting; α-, anti-. (a and b) Native or HA-tagged TSG101 or p53 proteins in Western blots were detected by anti-p53 monoclonal antibody (a; AB-1; 1:1000; the secondary antibody was HRP-conjugated goat anti-mouse IgG, diluted 1:1000) or by anti-HA antibody labeled with HRP (b; diluted 1:500). (c and d) Western blot detection of immunoprecipitated proteins analyzed with anti-MDM2 antibody.