Figure 1
Western blots for the HIV-1 gag proteins in HIV-1 produced in tissue culture following treatment with protease inhibitors. U87/CD4/CXCR4 cells were plated in 6 well plates at 80,000 cells/well and allowed to grow to confluence. The cells were infected with either HXB2 or RF/V82F/I84V (protease inhibitor resistant virus) and RT activity was monitored. On day 3 of culture, infected cells were treated with 20 nM LPV, and 1 ml of media was removed at 0, 4, 8, 24, and 72 hours post-drug treatment. The virus was pelleted, and the pellet was then lysed using sodium-dodecyl sulfate (SDS) lysis buffer and then run on a 10% SDS polyacrylamide gel. Following transfer to nylon membranes, blots were probed with the primary mouse anti-p24 antibody and the horseradish peroxidase-conjugated goat anti-mouseantiserum. Films were exposed following treatment with the ECL kit (panel A). Ratio of unprocessed p55gag to processed CA p24 over a 72 hour time course was determined by scanning the blots and quantifying the bands (panel B).