An NF-kB reporter assay system was designed using a pGL4-NFkB construct (Promega). Quadruplicate repeats of the NF-kB binding consensus sequence 59-GGGAATTTCC-39 were excised from a p-NF-kB-Luc (Gift from Dr. Heike Laman, University of Cambridge) vector using KpnI and HindIII and ligated into the pGL4.20 vector multiple cloning site using the same restriction sites. pGL4.20 was chosen as it is optimized for more efficient expression in mammalian cells, compared to previous generations of luciferase constructs, and contains fewer transcription factor consensus sequences such that background luciferase expression is reduced. HFFs were electroporated in batches of 6610 5 cells per cuvet (Amaxa) with 2 mg of pGL4-NF-kB, pooled together and aliquoted at 10 5 cells per well of a 24-well plate. After 48-hours recovery in supplemented media, the transfected HFFs were serum-starved for five days. HFFs were inoculated with 1000 particles/cell of DgH HSV-1 and were lysed at the indicated time points in 100 ml passive lysis buffer (Promega) and 20 ml of lysate were examined for luciferase activity using 50 ml assay buffer (Promega).