Figure 5
From: Constitutive degradation of IκBα in human T lymphocytes is mediated by calpain
Effect of E64D on stimulus-dependent modification of IκBα (Integrated Densities). T cells obtained from young and elderly donors were either left untreated or treated with E64D (50 μM) for 45 min. At the end of incubation cells were stimulated with 100 μM pervanadate or TNFα (10 ng/ml) for 10, 20 or 30 min. Cell lysates were prepared and resolved by SDS-PAGE, using equal amounts of protein (30 μg / lane). Resolved proteins were transferred to nitrocellulose membrane and Western blotted with antibody to IκBα and detected using ECL. Mean integrated densities of IκBα specific bands obtained from a minimum of 4 donor pairs were used to determine fold change when compared to their respective controls and are presented as Mean (fold-change) ± S.D. [TNF-α (A); Pervanadate (B)].