Figure 7

Cav-1 over-expression reduces the cholesterol content in HIV-1 virus particles. (A) Proviral DNA genome NL4-3 was transfected into U87 cells either with pCZ-vector or with pCZ-Cav-1. Virus particles generated were concentrated and normalized by p24 assay. Cholesterol contents were measured using the Amplex Red cholesterol Assay Kit. The results are presented as percentage of cholesterol content relative to control (HIV/pCZ-vector, set as 100%) and are the mean± SD of triplicate experiments with P values. (B) Normalized samples were replenished with exogenous cholesterol, and the level of infectivity was measured by infection of TZM-bl cells and luciferase assay. All experiments were performed in triplicate, and results shown are mean ± SD with P values. (C) Normalized samples from samples of (7A) were subjected to Western blot analysis using antibody to Gag, Nef, and Cav-1 to determine whether Cav-1 protein is incorporated in virus particles.