Figure 2

Adenoviral-mediated Expression of mIκBα Blocks NF-κB Transcriptional Activity. (A) BCPAP, SW1736, 8505C, TPC1, and C643 cells were transduced with Ad-mIκBα at an MOI of 0, 5, 25, 50, 100, and 250. After 48 hours, whole cell lysates were prepared, and expression levels of mIκBα were assessed by Western blot analysis. α-tubulin was used as a loading control. (B) Cells were transfected with an NF-κB-responsive luciferase reporter and actin-β-gal expression vector to control for transfection efficiency. Cell lysates were prepared 24 hours after transfection, after which luciferase and β-gal activities were assayed. Data is represented as NF-κB activity [luciferase activity/β-gal activity; relative light units (RLU)]. Mean ± S.E.M. of 3 independent experiments performed is reported. (C) Cells were transduced with Ad-mIκBα at an MOI of 0, 5, 25, 50, 100, and 250. After 24 hours, cells were then transfected with an NF-κB-responsive luciferase reporter and actin-β-gal expression vector to control for transfection efficiency. Cell lysates were prepared 24 hours after transfection, after which luciferase and β-gal activities were assayed. Data is represented as fold NF-κB activity where the basal activity (untransduced) of each cell line was normalized to 1. Mean ± S.E.M. of 3 independent experiments performed is reported.