Fig 6.

VEGF-A increases VV gene expression and replication in normal human bronchial epithelial (NHBE) cells, events that are blocked by inhibition of Akt. (A) NHBE cells were stimulated with rhVEGF (10 ng/ml) or mock medium for 10 min before infection with VVL15 at an MOI of 1 for 24 h. Bioluminescence was quantified using the IVIS imager. (B) NHBE cells were transiently exposed to 20 μM Akt inhibitor VIII (Akt-i) or DMSO for 2 h. Cells were then infected, with or without prior stimulation with rhVEGF (10 ng/ml) (10 min prior to infection), with wild-type VV at an MOI of 0.1. Viral replication was quantified at 72 h postinfection (p.i.) using a TCID₅₀ assay. The viral replication was presented as fold change normalized by the viral production in NHBE cells in normal culture medium. Each sample was assayed in triplicate, and each time point assay was performed in biological duplicates. An unpaired two-tailed Student t test was used to compare means. ***, P < 0.001.