Fig 5.

Infection with TCRV or rLCMV/NP* D382A does not prevent SeV-mediated induction of NF-κB-dependent transcriptional activation. A549 cells were mock infected or infected (MOI = 0.1) with TCRV or rLCMV/NP* D382A and at 72 h p.i. seeded on 24-well plates (2 × 10⁵ cells/well) prior cotransfection with pNF-κB-Fluc (500 ng) and pSV40-RL (50 ng) plasmids for 5 h, followed by infection with SeV (MOI = 3) for 18 h, at which time cell lysates were prepared to determine levels of Fluc (NF-κB activation) and RL (normalization of transfection efficiency). (A) Reporter gene activation is expressed as fold induction over the level seen with the empty vector-transfected and mock-infected control cells. (B and C) From duplicate wells, cells were fixed to determine percentages of antigen-positive cells by immunofluorescence using a mouse hyperimmune serum to TCRV or monoclonal antibody 1.1.3 to LCMV-NP (B), and tissue culture supernatants (TCS) were collected to determine the production of infectious virus (TCRV and rLCMV/NP* D382A) progeny (FFU per milliliter) using a focus-forming-unit assay (C). Values shown correspond to averages ± SD of the results from two of three independent experiments.