Fig. 2.

Effect of Cif on NF-κB activity upon stimulation with TNFα or infection with B. thailandensis. (A) Cells transfected with the indicated expression plasmids and an IL8 luciferase reporter plasmid were treated with 30 ng/ml TNFα and 1 μM MLN4924 for 8 h. NF-κB activation was then measured and fold change was presented as mean±s.d. The results shown are representative of two independent experiments. (B) Cell lysates from the reporter assay described in Fig. 3A were resolved using SDS-PAGE and the cellular protein level of phosphorylated IκBα was determined using Western blotting. (C,D) Cells transfected with the indicated expression plasmids and an IL8 luciferase reporter plasmid were pre-treated with 1 μM MLN4924 for 1 h prior to infection with B. thailandensis for 15 h as described in the Materials and Methods. NF-κB activation was then measured at the stipulated time and fold change was presented as mean±s.d. The results shown are representative of two independent experiments.