Figure 10. ICP0Δ and vIET mutant virus replication. (i).

GPL cells transiently transfected with gpPML (+) before being infected with ICP0Δ or IET mutant virus, WT HSV, or GPCMV (0.1 MOI). (−) indicates GPL cells with intrinsic gpPML only. Cell monolayers and supernatant were harvested 3 days p.i. and titrated on GPL cells. Results plotted as virus titer versus experimental condition. (ii) IFN-I susceptibility of vIET mutant compared to WT. Cells were incubated overnight in the absence or presence of 100 IU/ml (red) or 1000 IU/ml (green) human IFN-I prior to virus infection (0.1 MOI). After 72 hrs, supernatant and monolayer were harvested and titrated in duplicate on GPL cells. Results blotted as percentage of virus growth. (iii) GPL cells were grown in the absence (−) or presence (+) of ruxolitinib (10 μM) and IFN-I (100 IU/ml). Cells were pretreated 6 hrs prior to infection with vIET (0.1 MOI). Cells and supernatants were harvested 4 days p.i. and titrated on GPL cells. Results plotted as virus titer versus experimental condition. (iv) Complementation of ICP0Δ mutant virus by GPCMV. GPL cells were infected with either WT GPCMV (yellow) or vIET (green) (3×10⁴ pfu) prior to being superinfected with ICP0Δ mutant virus (10² pfu). ICP0Δ mutant virus alone (red) served as control. Cells and supernatants were harvested after 3 days and titrated on U2OS cells. Results plotted as virus titer versus experimental condition.