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Fig. 4. Inhibition of either tyrosine or serine I{kappa}B{alpha} phosphorylation stimulates apoptosis in an injury-context specific fashion. HeLa cells were infected with Ad.BglII, Ad.I{kappa}B{alpha}(S32/36A) or Ad.I{kappa}B{alpha}(Y42F) at an moi of 1000 particles/cell for 24 hours. Cells were treated with UV (50 J/m2), TNF-{alpha} (10 ng/ml), pervanadate (50 µM) or H/R (5 hours hypoxia, 18 hours reoxygenation) and then harvested 18 hours after treatment. Cells were then stained with annexin-V—FITC and propidium iodine. Results depict the mean (±s.e.m., n=3) percent of apoptotic cells as shown by FACS analysis. Ad.I{kappa}B{alpha}(S32/36A) more significantly increased apoptosis following UV or TNF-{alpha} treatment, whereas Ad.I{kappa}B{alpha}(Y42F) preferentially increased apoptosis following pervanadate or H/R treatment. Paired t-test analysis was performed between Ad.BglII infected and Ad.I{kappa}B{alpha}(S32/36A) or Ad.I{kappa}B{alpha}(Y42F) infected samples for each stimulus, and a statistically significant difference is denoted by *(P<0.05) or {dagger}(P<0.001). P-values for paired t-test analysis comparing Ad.I{kappa}B{alpha}(S32/36A) and Ad.I{kappa}B{alpha}(Y42F) infected samples for each stimulus are denoted above brackets.





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