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Fig. 7. Effects of TRIP6 RNAi on NF-{kappa}B activation by various stimuli. (A) TRIP6 RNAi inhibits the expression of transfected TRIP6. 293 cells (2x105) were transfected with 1 µg of an expression plasmid for Flag-TRIP6, 1 µg of an expression plasmid for Flag-IKK{alpha} and the indicated amounts of TRIP6 RNAi plasmid (pSuper-TRIP6). 14 hours after transfection, cell lysates were analysed by western blot with anti-Flag antibody. (B) TRIP6 RNAi inhibits expression of endogenous TRIP6. 293 cells were transfected with pSuper or pSuper-TRIP6. Transfected cells were selected with puromycin (10 µg ml-1) for 1 week. Cell lysates were analysed by western blot with a rabbit anti-TRIP6 and a monoclonal anti-ß-actin antibody. (C) Effects of TRIP6 RNAi on NF-{kappa}B activation by various stimuli. NF-{kappa}B reporter gene assays with the cell lines stably transfected with TRIP6 RNAi or control vector were performed as in Figs 4, 6.