Figure 8.

p65RelA interaction with Importin-α3 is competitively inhibited by KPV. A. Representative dot blot experiment. BL21 E. coli transformed with Imp-α3 were induced for 6h with 0.2mM IPTG at 25°C. Cells were lysed and then equal concentrations of lysate protein were spotted onto nitrocellulose. A bacterial lysate from identically treated BL21 cells which had not been transformed with the Imp-α3 vector served as a negative control. Dots were incubated in tris buffered saline containing 10μg.ml^-1 protein of a lysate from HBE cells expressing pCMV-FLAG-p65RelA and KPV at the concentrations shown. Dots were visualised using Anti-FLAGHRP. B. Competitive ELISA assay for the p65RelA:Imp-α3 interaction. Open circles: Control (plate coating = bacterial lysate not expressing Imp-α3); Closed Circles: Imp-α3-p65RelA interaction (plate coating = Imp-α3). Values are mean ± SEM, n=10. *P<0.05.