Adenovirus-mediated IκBΔN expression abrogates NF-κB activity and affects growth of L428 cells.
(A) Expression of IκBΔN in L428 cells. L428 cells were infected with an adenovirus expressing IκBΔN or with a control adenovirus containing an empty expression cassette (Ad5-IκBΔN and Ad5-control; MOI 300). Whole-cell extracts of infected and control cells (MCF-7, Reh) were prepared at the indicated time points and analyzed by Western blotting with anti-IκBα antibody. Note that L428 cells are defective in wild-type IκBα expression. (B) NF-κB binding activity. Whole-cell extracts of L428 cells infected with Ad5-control and Ad5-IκBΔN were analyzed by EMSA using H2K and H2B binding site probes for NF-κB (upper panel) and Oct-1 (lower panel), respectively. (C) Growth rate. Numbers of noninfected or infected L428 cells (as indicated) were determined for 3 days starting with 5 × 105 cells. Five independent experiments were performed. NI indicates noninfected; ND, not determined.
