Figure 2

Inactivation of Foxo3a by LMP1 results in Id1 induction. (A) NP69 cells stably expressing pLNSX vector control or LMP1-pLNSX were transfected with a HA-tagged Foxo3a expression vector. Forty-eight hours after transfection, cells were cultured in growth factor free medium for 6 hrs prior to immunoblotting analysis. (B) HEK-293 cells were transfected with an Id1 (-1695) promoter luciferase reporter and various doses of LMP1 expression vector (pSG5-LMP1) or (C) various doses of Foxo3a expression vector together with 40 ng LMP1 expression vector or control pSG5 empty vector. (D) NP69 cells were transfected with an Id1 (-353) promoter luciferase reporter and various doses of a LMP1 expression vector (pSG5-LMP1) or (E) various doses of a Foxo3a expression vector together with 40 ng LMP1 expression vector or pSG5 control vector. Thirty hours post-transfection, cells were incubated in serum free medium for 6 hrs prior to harvesting for luciferase analysis.