Figure 5

Role of Vpr oligomerization in incorporation of Vpr into virus particles. (A) Visualization of Vpr^wt and Gag interaction in HeLa cells by BiFC. HeLa cells were transfected with VN-Vpr^wt and VC-Gag expression plasmids. Thirty-six hours post transfection, cells were fixed, stained with DAPI and analyzed for presence and pattern of BiFC signal at 60× magnification. (B) Incorporation of Vpr mutant molecules into virus particles. Vpr plasmids were cotransfected with pNL43ΔVpr proviral plasmid in HEK293T cells. Forty-eight hours post-transfection, supernatant was collected, filtered, lysed and subjected to SDS-PAGE electrophoresis and evaluated for the presence viral proteins p24-Gag and Vpr using p24-specific and Vpr specific antibodies by western blot. Figure represents one of five independent experiments (n = 5) with similar results.