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. 2010 Dec 23;22(10):1225–1237. doi: 10.1089/hum.2010.012

FIG. 3.

FIG. 3.

Intracellular localization and virion incorporation of different R88-A3G mutants. (A) HeLa cells were transfected with equal amounts of HA-A3G or R88-A3Gwt/mutant plasmid DNA. Forty-eight hours post-transfection, cells were fixed and treated with rabbit anti-A3G polyclonal antibody followed by incubation with fluorescein isothiocyanate (FITC)-conjugated anti-rabbit antibody and observed by fluorescence microscopy ( × 20 objective). (B) 293T cells were transfected with HxBru-Vif+ (3 μg; lane 2) or co-transfected with HxBru-Vif+ and 2 μg of R88-A3Gwt/mutant (D128K, P129A, or E259Q; lanes 3 to 6). After 48 hr, the produced virus particles were collected from the supernatant by ultracentrifugation through a 20% sucrose cushion. The virus lysate samples were loaded onto a 12% SDS-PAGE gel and analyzed by Western blotting with rabbit anti-A3G and anti-p24 antibodies, as indicated. The result shown is representative of two independent experiments.