Construction of a recombinant adenovirus for expression of dnIKK2 (AdV-dnIKK2).

(A) A dominant-negative form of IKK2 was generated by mutating K44 to A, and a Flag tag engineered to its 5′ end. The construct was cloned into the adenovirus transfer vector pACCMVpLpASR and cotransfected together with pJM17 into 293 cells.22 (B) Western blot analysis of Flag-dnIKK2 expression in 293 cells. Cells were incubated with deceasing amounts of plaque-purified AdV-dnIKK2 (lanes 1-3; lane 4: no virus). The Flag-dnIKK2–specific band is indicated by an arrow. ns indicates nonspecific.