Figure 7
Comparative analysis of the GFP-related fluorescence associated with NefG3C-GFP and GFP-Vpr VLPs. (A) The indicated amounts of NefG3C-GFP (b) and GFP-Vpr (c) VLPs purified from the supernatants of transfected 293T cells were incubated with aldehyde latex beads and then analyzed by FACs. As control, the analysis of beads incubated with equivalent amounts of void VLPs (a) was included. The mean fluorescence values for each slope are indicated in the respective panels. The results are representative of data obtained from two independent experiments. (B) Analysis of the GFP-related fluorescence associated with MLV-based VLPs produced in cells expressing NefG3C-GFP (b) or wt Nef-GFP (c). 293T cells were co-transfected with a MLV packaging construct and a vector expressing wt Nef-GFP or NefG3C-GFP. VLPs released in the supernatants were purified, and the relative amounts quantified through the Mn++ dependent reverse transcriptase assay. Amounts of VLPs equivalent to 106 or 5 × 105 cpm were incubated with aldehyde latex beads and analyzed by FACs as compared to equal amounts of void MLV VLPs (a). The mean fluorescence values for each slope are indicated in the respective panels. The results are representative of data obtained from two independent experiments.