Disruption of both Pr55Gag zinc fingers affects VLP production. (A) Schematic representation of the Gag fusion proteins used in the BRET and release assays. (B) Wild-type or mutated YFP-fused Gag proteins were expressed in 293T cells for twenty-four hours. VLPs in the cell supernatant were purified. Cell lysates and VLPs were analyzed by Western blotting using anti (α)-GFP antibodies. Anti (α)-GAPDH antibodies were used as loading controls. This figure is representative of three independent experiments. (C) 293T cells were transfected with constant amounts of pCMV-Pr55Gag-Rluc and increasing amounts of wild type or mutated YFP-fused Gag expressors. Twenty-four hours post-transfection, cells were collected and BRET ratios determined. BRET ratios are plotted in function of their corresponding total YFP/Rluc ratio. This figure is representative of three independent experiments.