FIG 4.
Compound 1 does not markedly affect processing of Pr55Gag during particle maturation. (A) Immunoblot analysis of HIV-1 particles produced in the presence of the indicated concentrations of C1. Pelleted particles were lysed and fractionated by SDS-PAGE, and proteins were detected by immunoblotting using a monoclonal antibody to HIV-1 CA (left) and a mixture of anti-CA and anti-Env (gp41 + gp120) antibodies. (B) Pulse-chase analysis of HIV-1 processing during particle maturation. HEK293T cells producing HIV-1 particles were cultured in the presence or absence of C1 (100 μM) and radiolabeled with [35S]methionine plus cysteine for 30 min, followed by culture in the presence of unlabeled medium. Supernatants and cells were collected at the indicated times of chase, and protein extracts were immunoprecipitated using a CA-specific monoclonal antibody and separated by SDS-PAGE. Bands were detected by phosphorimager analysis. The images shown in panels A and B are representative of results obtained from two and three independent experiments, respectively.