Figure 4. Cleavage of PABP in vitro by purified recombinant HIV-1, HIV-2 and MoMLV proteases.
Crude HeLa S10 extracts (50 μg) were incubated with 3 ng/μl of each recombinant protease in a total volume of 20 μl for 90 min and subjected to Western blotting analysis by using: (A) a monoclonal antibody against PABP (10% gel); (B) a polyclonal antiserum against the 100 kDa subunit of eIF3 (top panel; 10% gel), an antibody against eIF4A (middle panel; 15% gel) or a monoclonal antibody raised against eIF4E (bottom panel; 15% gel). MW, molecular mass (sizes given in kDa).