Figure 5

Over-expression of Akt and MEK enhances RV-induced apoptosis. RK13 cells were transfected with eukaryotic expression vector pUSEamp(+) containing constitutively active HA-tagged MEK1 or myristoylated myc-tagged Akt1 under the control of a CMV promoter, or with an empty pUSEamp(+) control. A – Expression of MEK1 and Akt1 was determined by Western blotting. Cell lysates were collected 24 hours post-transfection and 30 μg protein separated by SDS-PAGE and transferred to nitrocellulose membranes. MEK1 and Akt1 were detected by anti-HA and anti-myc antibodies respectively. B – RK13 cells in 6-well plates were transfected with Akt, MEK or pUSEamp(+) control constructs for 24 hours and subsequently infected with RV or mock-infected. 24 hours later cell lysates were collected and tested for caspase activity using artificial caspase substrate Ac-DEVD-pNA.