Fig. 7: SIRT1 and SIRT2 remove methacryl from Kmea.

a Acid extracted HeLa histones were incubated with or without recombinant HDAC3 for 12 h at 37 °C. Samples were then analyzed by western blot. b HEK 293 T cells were treated for 24 h with either 10 mM sodium butyrate or 1 μM TSA. The core histones were prepared by acid extraction method and then subjected to western blot analysis. c, d Quantitation of deacylase activity of SIRT 1-7 for in vitro screen using either H3K18mea peptide (c) or H3K18ac peptide (d) is shown. The deacylase activity percentage was calculated by dividing the area of the H3K18 unmodified chromatogram peak over the sum of the H3K18 unmodified and modified chromatogram peaks. e H3K18mea synthetic peptides were incubated with SIRT1 or SIRT2 in presence of reaction buffer with or without its cofactor NAD + . Chromatograms of each sample condition are shown. Open diamonds indicate the H3K18 unmodified peptide peaks. Filled diamonds indicate the H3K18mea peptide peaks. f Acid-extracted HeLa histones were incubated with or without recombinant SIRT1 or SIRT2 for 12 h at 37 °C. Samples were then analyzed by western blot. g HEK 293 T cells were transfected with either control empty pcDNA 3.1 vector or pcDNA 3.1 SIRT2-Flag. Whole cell lysates were prepared 48 h post-transfection and then subjected to western blot analysis with the indicated antibodies.