FIG. 1.

Generation of CD40L-pseudotyped SIVs. (A) CD40L-dSIV was generated by transient cotranfection of pCD40L, pVSV-G, and pSIVΔEΔNgfp into 293T cells. pSIVΔEΔNgfp is a plasmid encoding the full-length SIVmac239 proviral DNA with deletions in the env and nef genes and an insertion of the GFP gene in the nef region. pVSV-G is the plasmid containing the VSV-G gene under a CMV promoter. pCD40L contains the CD40L gene under a CMV promoter. Deletions are indicated by shaded regions, the inserted GFP gene is shown as an open box, and pA refers to the polyadenylation signals from the expression plasmids. (B) 293T cells cotransfected with pCD40L, pVSV-G, and pSIVΔEΔNgfp were able to express all three genes (CD40L, VSV-G, and GFP). Cells were fixed, permeabilized, and double stained with anti-CD40L conjugated with Alexa Fluor 555 and anti-VSV-G conjugated with Alexa Fluor 647. Protein expression was visualized by confocal microscopy.