Fig 5. Virus replication and gB expression in infected mice.
(A) Virus replication in tears of infected mice. WT, M2-/-, and M2-OE mice were ocularly infected with 2 X 105 PFU/eye of McKrae virus without corneal scarification. Tear films were collected on days 1–7 PI and virus titers were determined by standard plaque assays as described in Materials and Methods. Each point represents the mean ± SEM of 20 eyes per group. (B) Expression of gB in corneas of infected mice. WT, M2-/-, and M2-OE mice were infected ocularly as above. gB expression in corneas of infected mice was determined on day 4 PI. In each experiment, the estimated relative copy number of HSV-1 gB was calculated using standard curves generated from pAC-gB1. Briefly, DNA template was serially diluted 10-fold such that 5 μl contained from 103 to 1011 copies of LAT, then subjected to TaqMan RT-PCR with the same set of primers. By comparing the normalized threshold cycle of each sample to the threshold cycle of the standard, the copy number for each reaction was determined. GAPDH expression was used to normalize the relative expression of each transcript in corneas and TG of infected mice. Each bar represents the mean ± SEM from 3 mouse corneas. Panels: A) Virus titer; and B) gB expression.