Fig. 1. Ad does not impair Mnk1 kinase activity. 293 cells were transfected with plasmids expressing GST–Mnk1 or GST–T2A2, a kinase-deficient mutant of Mnk1 (Waskiewicz et al., 1997). Cells were either uninfected (uninf.), treated with 30 ng/ml EGF for 15 min (Feigenblum et al., 1998), or infected with wtAd for 36 h (late Ad inf.). GST–Mnk1 or GST–T2A2 was recovered from equal amounts of cell lysates by glutathione–Sepharose chromatography and incubated with 1 µg of purified recombinant eIF4E and [γ-³²P]ATP, resolved by SDS–PAGE and autoradiographed (top panel), or subjected to immunoblot analysis with anti-GST antibodies (middle panel). eIF4E levels are shown by immunoblot (bottom panel). Quantitation was performed by digital densitometry.