Virus-producing constructs used to make pseudotyped lentiviruses.

<p>(A) Schematic diagrams of constructs encoding the lentiviral backbone FUGW and envelope glycoproteins. Ubi: the human ubiquitin-C promoter; GFP: enhanced green fluorescence protein; WRE: the woodchuck hepatitis virus posttranscriptional regulatory element (WRE) to increase the level of transcription; ΔU3: deleted U3 region that results in the transcriptional activation of the integrated viral LTR promoter; pA: polyadenylation signal; E1, E2, 6k, E3: SFV-glycoprotein (E1 for fusion, E2 for receptor binding, 6k a linker, and E3 is a signal sequence). The VSV-G expressing plasmid contains the rabbit β-globin intron and poly(A) signal. (B) Viral supernatants harvested from virus-producing cells transiently transfected with GFP-vpr, SFV-G, or VSV-G, and other necessary packaging constructs, were coated to a poly-lysine containing coverslip by centrifugation. The resulting coverslips were then rinsed and immunostained with an anti-SFV-G antibody (red) to label the glycoproteins and imaged using a laser confocal microscope.