(A) Endogenous SIRT1 and BMAL1 expression in MEFs after serum shock were determined by western blot.
(B and C) Sirt1 (B) and Dbp (C) genes expression in liver was analyzed by quantitative PCR. The highest expression time point was set to 1.
(D) Endogenous SIRT1 proteins from livers of entrained mice at indicated time points were immunoprecipitated and subjected to deacetylation assays. The value at ZT15 was set to 1. Top panel shows the amount of immunoprecipitated SIRT1.
(E) Cell extracts prepared from serum-shocked MEFs at indicated time points were complemented with recombinant SIRT1 protein and acetylated p53 peptide as substrate and subjected to deacetylation assay. Value at time 0 was set to 1.
(F) Liver extracts prepared from entrained mice were complemented with recombinant SIRT1 protein and acetylated p53 peptide as substrate and subjected to deacetylation assay. Values at ZT15 were set to 1. All data presented are the means ± standard errors of the mean (SEM) of three independent samples.