FIG. 5.
Detailed examination of Gag-Pol and Gag release from actinomycin D- and leptomycin B-treated cells. Relative RT measurements were normalized for cell viability to correct for drug-induced cytotoxicity (A). Aliquots of HIV-1- and HIV-2-infected cells were examined at each time point to assess fraction of viable cells. Levels of Gag were assayed by Western blotting (B). Time courses were performed with actinomycin D (5 μg/ml) or leptomycin B (10 nM for HIV-1; 20 nM for HIV-2). The HIV-1 antibody is reactive against p24CA, and the HIV-2 antibody (Chemicon) is reactive against p26CA; both also react with capsid-containing Gag and Gag-Pol products. Act D, actinomycin D; LMB, leptomycin B; M, mock-infected cells.