Fig 4.
Design of ICP47 promoters and HSV-1 recombinants to investigate ICP47 promoter activity. (A) Schematic showing the features of the region upstream of the ICP47 open reading frame in the native locus (top) and as used in recombinant viruses, where it drives Cre from between UL3 and UL4. Deletions are shown with dotted lines and single bp changes are indicated with asterisks. (B and C) Replication of HSV-1 recombinants as marked in vitro (B) and in vivo (C), markers and symbols and statistical analyses are the same as in previous figures. (D) Promoter activity was measured based on GFP expression for recombinant viruses under control (no drug) or cycloheximide release conditions (CHX/AD) from the viruses as shown. (E–H) Groups of ROSA26 mice were infected with HSV-1 pICP47_eGC_full (E and F) or pICP47_eGC_mut (G and H) and DRG were taken and stained for β-gal expression at the days shown. The number of β-gal+ cells per mouse (E and G) and the number of DRG per mouse (F and H) are shown as in the previous figures. Data are pooled from three independent experiments with at least two overlapping time points. Differences between means were determined using a one-way ANOVA with Bonferroni’s multiple-comparison test (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; and red stars signify differences in latency).