Fig. 6

Chimeric UL52–UL54 mRNA is expressed with late kinetics and by multiple HSV-1 strains. a The UL52–UL54 fusion transcript. b Assessment of the UL52–UL54 splice junction usage at different times after infection by real-time RT-qPCR. Increased RNA abundance is reflected as lower crossover threshold (Ct) values and normalized to 18S rRNA. Three technical replicates were utilized per condition/time point. Representative data are shown from one of three biological replicates. c Detection of the unique UL52–UL54 splice junction by RT-PCR using a primer scanning the splice junction. NHDFs were infected in parallel with either HSV-1 strain Patton (lanes 2–7) or with wild-type strain 17 syn+ (lane 9), KOS (lane 10), strain F (lane 11) viruses, or with n12, a KOS ICP4 null mutant (lane 12), and RNA was collected at either 6 h (lane 4) or 18 h (lanes 2–3, 5–7, and 8–12) post infection. Inhibitors of protein synthesis (cycloheximide, CHX) or the viral DNA polymerase (phosphonoacetic acid, PAA) were included as indicated (lanes 4–6). Amplification products were visualized with ethidium bromide