Figure 3. Generation of Single Cycle Infectious Virions Containing HERV-K_CON Genomes and Gag-PR-Pol.

(A) Schematic representation of the HERV-K_CON–based CHKCG genome. Changes to HERV-K_CON are depicted in white boxes. The 5′ LTR was modified to include the CMV promoter inserted in place of the U3 region. The Env ORF, which now contains CMV-GFP, is disrupted.

(B and C) Photomicrographs of 293T cell monolayers after infection with CHKCG-carrying HERV-K_CON(VSV-G) pseudotyped virions, which were generated following transfection of 293T cells with the indicated plasmid mixtures.

(D and E) Infectious titers of HERV-K_CON(VSV-G) pseudotyped virions generated following transfection with the indicated plasmid mixtures in the absence (D) or presence (E) of K-Rev/Rec and using 293T target cells. GFP-positive foci were enumerated visually and expressed as infectious units per milliliter of virion-containing supernatant.

(F) Infectious titers of CHKCG containing HERV-K_CON(VSV-G) using 293T target cells in the presence or absence of 50 μM AZT.

All data are representative of at least three experiments.