Hydrophobic residues on Vpr helix-1 are important for incorporation into virions.

<p>(<b>A</b>) 293T cells were co-transfected with pcDNA3-hVpr plasmids expressing WT or mutant Vpr (or the empty vector control), pLVSV-G, and pNL4-3_{e-n-GFP RTm, VprΔ22-86} to produce virus for virion delivery of Vpr. At day 2 the virus stocks were harvested and the 293T cells were lysed. Western blot indicates protein levels for WT or mutant Vpr (bottom). A western blot for β-actin is used as a loading control (top). (<b>B</b>) Lysates were prepared from the virus stocks in (A). Viral lysates were analyzed for protein levels of WT or mutant Vpr by western blotting as in (A) (bottom). The HIV-1 p24 capsid (p24 CA) is shown as a protein loading control (top). (<b>C</b>) Densitometry of all bands in (B) was performed. The intensity of the Vpr protein band was normalized to p24 CA and plotted as % Vpr incorporation relative to WT Vpr for each mutant. The data are shown as the mean ± the standard deviation of three independent experiments.