Universal human ifn 1

On day 0, GPL cells were pre-treated with 100 IU/ml or 1000 IU/ml universal human IFN-I (PBL Assay Science). The next day, IFN pre-treated and control untreated cells were infected separately with either WT GPCMV or vIET mutant virus (0.1 MOI) and incubated at 37°C with 5% CO₂. After 72 hrs, supernatants and monolayers were harvested and titrated in duplicate on GPL cells as previously described. IFN-I IU/ml were confirmed by VSV growth experiments in the presence or absence of different concentrations of IFN-I on GPL cells prior to reported studies (data not shown). GPL cells were pre-treated with 10 μM of the JAK1/JAK2 inhibitor ruxolitinib (Invivogen) for 1–6 hrs. Depending on the experimental condition, some cells were treated with 100 IU/ml of universal human IFN-I (PBL Assay Science). Cells were subsequently infected with vIET (0.1 MOI) for 1 hr. Cells were washed and fresh F-12 medium supplemented with 10% FCS plus 10 μM ruxolitinib were added then incubated at 37°C with 5% CO₂ for 4 days. Cells and supernatant were harvested and virus titrated on GPL cells.