Fig. 3. IB(Y42F) inhibits NFB activation following pervanadate and H/R treatment. (A) HeLa cells were co-infected with Ad.IB(Y42F) or Ad.BglII together with Ad.NFBLuc 24 hours before treatment. Cells were treated with UV (50 J/m²), TNF-(10 ng/ml), pervanadate (50 µM) and H/R (5 hours hypoxia, 6 hours reoxygenation) then harvested 6 hours after treatment. Whole cell extracts were normalized by total protein content and subjected to luciferase assay. NFB activity was determined by the relative luciferase activity (as light units). The relative NFB activation was calculated by deducting the mean NFB baseline activation for each vector group (in the absence of stimulation) from the corresponding individual values from the same vector group in the presence of stimulus. The relative NFB activation is plotted for each individual stimulus (±s.e.m., n=6). (B) Percent NFB inhibition by IB(Y42F) for each experimental point was calculated using the following formula: percent inhibition=1—(relative activation of each Ad.IB(Y42F) infected sample/mean relative activation of the Ad.BglII infected group). Results depict the mean (±s.e.m.) for n=6 independent data points in each group.

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