FIG 7.
Effects of PF-46396 analogues on virus release efficiency of the SP1-A3T mutant. HeLa cells were transfected with pNL4-3/WT or pNL4-3/SP1-A3T and cultured with 5 μM compound or an equivalent volume of DMSO. Cells were metabolically labeled for 2 h with [35S]Met-Cys, and released virions were pelleted by ultracentrifugation. (A) Cell and virus lysates were immunoprecipitated with HIV-Ig and analyzed by SDS-PAGE and fluorography. (B) Phosphorimager analysis was used to quantify virus release efficiency, calculated as the amount of particle-associated Gag as a fraction of total (cell plus virion) Gag. Statistical significance was assessed by using one-way analysis of variance followed by Dunnett's multiple-comparison posttest to compare SP1-A3T virus release efficiency in the presence of DMSO to that in the presence of each test compound. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Mean values from independent experiments are presented (n = 2 to 4), and error bars indicate standard deviations. Statistical analysis was performed by using GraphPad Prism version 6 software. The compound's ability to inhibit HIV-1 is indicated (A, active; I, intermediate activity; N, nonactive).