Fig 5.
Transient transfection of VHS suppresses the expression of phospho-PKR (p-PKR) in HT1080 cell lines. Cells were transfected with 1.5 μg of plasmid DNA expressing wild-type (pVHS) or catalytically inactive (pVHSm) VHS protein under the control of the CMV promoter either alone or in the presence of plasmids expressing two other tegument proteins, UL48 (pVP16) and UL49 (pVP22). Plasmid pMTS1 was used as a control. The transfected cells were incubated for 48 h before being harvested for detection of total PKR and phosphorylated forms of PKR on threonine 446. The fold induction of both the phosphorylated form of PKR and PKR were determined separately using the TINA program to measure the intensity of the band normalized to GAPDH.