FIG 11.
The inactive analogue StA-MAT-23 interferes with PF-46396 activity. HeLa cells were transfected with pNL4-3/WT and cultured with the indicated concentrations of PF-46396 and/or StA-MAT-23. Cells were metabolically labeled for 2 h with [35S]Met-Cys, and released virions were pelleted by ultracentrifugation. (A) Virus lysates were immunoprecipitated with HIV-Ig, and processing of CA-SP1 to CA was analyzed by SDS-PAGE and fluorography. (B and C) Phosphorimager analysis was performed to quantify the percentage of CA-SP1 relative to total CA-SP1 plus CA in pelleted virus samples. The data are plotted as the percentage of CA-SP1 in the presence of increasing concentrations of PF-46396 (B) or StA-MAT-23 (C). (D) The percentage of CA-SP1 in the presence of 1 μM PF-46396 was compared to the percentage of CA-SP1 in the presence of 1 μM PF-46396 plus increasing concentrations of StA-MAT-23. Statistical significance was assessed by using one-way analysis of variance with Dunnett's multiple-comparison posttest. *, P < 0.05. (E) The percentage of CA-SP1 in the presence of increasing concentrations of PF-46396 was compared with that in the presence of the equivalent concentration of PF-46396 plus 1 μM StA-MAT-23. Statistical significance was assessed by using one-way analysis of variance with Bonferroni's multiple-comparison posttest. *, P < 0.05; **, P < 0.01. Mean values from independent experiments are presented (n = 3), and error bars indicate standard deviations. Statistical analysis was preformed by using GraphPad Prism version 6 software.