FIG. 7.

Fate of the capsid in HeLa cells infected with HIV-1 CA variants. (A) VSV-G-pseudotyped (Env+) wild-type (wt) or A92E mutant HIV-1 virions were incubated with HeLa cells in the presence or absence of 5 μM Cs. As a control, virions lacking envelope glycoproteins (Env−) were also incubated with HeLa cells under the same conditions. At 4 h after infection, the cells were washed and incubated at 37°C in fresh medium for another 12 h. After lysis of the target cells, cytoplasmic lysates were analyzed directly (“Input”) or were separated over a 50% sucrose cushion into supernatant (“Sup”) and particulate (“Pellet”) fractions. The fractions were analyzed by Western blotting, with a monoclonal anti-HIV-1 p24 antibody. The experiment was repeated twice with similar results. (B) HIV-1 CA mutants were used in the same experiment as that described in panel A, in the absence of Cs. (C) HIV-1 CA mutants were used in the same experiment as that described in panel A, in the presence of 5 μM Cs. The results of two independent experiments (Exp 1 and 2) are shown in B and C. wt, wild type.