Bimolecular Complementation to Visualize Filovirus VP40-Host Complexes in Live Mammalian Cells: Toward the Identification of Budding Inhibitors
Figure 4
Inhibition of VP40-Tsg101 interactions and VLP budding. (a) Chemical structure of compound 5539-0062. (b) Budding assay and western blotting for eVP40 and mVP40 in VLPs and cell extracts from human 293T cells treated with DMSO alone, or the indicated concentrations of PTAP-inhibitor 5539-0062. (c) Bar graph of the relative amounts of eVP40-WT in cells and VLPs (part B) determined using Dot-Blot, LabWorks software (UVP). (d) Bar graph of the relative amounts of mVP40-WT in cells and VLPs (part B) determined using Dot-Blot, LabWorks software (UVP). (e) BiMC of human 293T cells expressing NYFP-Tsg101 + CYFP-eVP40-WT and treated with DMSO alone, or the indicated concentrations of PTAP-inhibitor 5539-0062. Cells were examined for YFP fluorescence at 24 hour p.t., and positive cells were quantified by FACS analysis. The average values were as follows: DMSO alone = 100%, 25 μM = 84%, 50 μM = 68%, and 75 μM = 24%.
