Figure 4. EVI1 negatively regulates bacteria-induced NF-κB activation via direct interaction with p65.
(A & B) A549 cells co-transfected with Flag-EVI1 and p65 were treated with NTHi for the times indicated in the figure, and the proteins were then immunoprecipitated with anti-Flag antibody (A) or anti-p65 antibody (B). Immunoprecipitations were then analyzed by western blot analysis with the indicated antibodies. (C) A549 cells co-transfected with Flag-EVI1 and p65 were treated with NTHi and then triple-immunostained with DAPI, anti-p65 and anti-Flag antibodies. Slides were visualized by fluorescence microscopy. (D) A549 cells were treated with NTHi or TNF-α for the times indicated in the figure, and the proteins were then immunoprecipitated with anti-p65 antibody and analyzed by western blot analysis with the indicated antibodies. Data are representative of at least three independent experiments.