Cell type: HeLa + Serum starvation (12 h) + SBA (5 mM, 24 h) Cross-linking conditions: No cross-linking Amount of chromatin per IP: 5×106 cells Amount of Ab per IP: 4 μg Beads type and amount per IP: 50 μL of Protein A/G MagBeads Comment: The ChIP was performed with 1 μg of normal rabbit IgG as a negative control. Real time quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human GAPDH CDS region, RPL30 Exon 3, FOXO3a-promoter and FOXO3a-downstream. Data are presented as enrichment of each sample relative to total amount of input chromatin at each amplicon.

Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:2000 Primary ab incubation condition: 2 hours at room temperature Secondary ab: Goat Anti-Mouse IgG H&L (HRP) Lysate: (-) HeLa, (+) HeLa+Sodium butyrate (30mM, 4hr) Protein loading quantity: 20 μg Exposure time: 60 s Predicted MW: 14 kDa Observed MW: 14 kDa