Figure 5. Primary T cells were isolated from WT and Sirt1^-/- mice and cultivated with anti-CD3 (5 µg/ml) and anti-CD28 (2 µg/ml) for overnight.

Cells were then treated with Res (10µg/ml) for additional 8 hours. (a) c-Jun acetylation in the lysates of treated cells was determined by immunoprecipitation with anti-c-Jun antibody and western blotting with anti-acetyl-lysine Abs (top panel). The expression levels of c-Jun (middle panel) and Sirt1 (bottom panel) in the whole cell lysate were confirmed by western blotting. (b) The levels of acetylated c-Jun were quantified and normalized with total c-Jun and its relative levels are shown. Error bars represent data from three independent experiments (mean + SD).