Figure 3. UL12.5 is expressed and functions during Phase I of HSV-1 reactivation.

3A-B. Neurons were latently infected with HSV-1 Stayput-GFP and reactivated in the presence of cycloheximide (CHX; 10 μg/ml) added at either the same time as LY294002 or 10 hours later. 3A. Relative expression of ICP27 mRNA quantified by RT-qPCR at 18 hours post-reactivation (Phase I).3B. Relative expression of gC mRNA quantified by RT-qPCR at 18 hours post-reactivation (Phase I).3C. Quantification of mtDNA abundance by qPCR of mtDLoop1 at 18 hours post-reactivation (Phase I). 3D. Relative expression of ICP27 mRNA, measured by RT-qPCR at 18 hours post-reactivation. Neurons latently infected with either KOS-SPA or KOS-UL98 were transduced with lentivirus expressing either GFP or UL12.5.3E. Representative immunofluorescence images (TIFs) of UL12/UL12.5 and TOM20 following reactivation. Dataset for histograms from two independent experiments representing the signal intensity distribution for anti-UL12/12.5 - BWp12 (Green) and TOM20 (Red). 3F. Quantification of UL12.5 expressing neurons as a percentage of neurons positive for UL12/UL12.5 staining per field of view at 0-, 4-, and 8 hours after reactivation. Each data point represents a field of view from two independent biological replicates.