Figure 3. SIRT6 deacetylates lysine 9 of histone H3 at telomeric chromatin.
a, Summary of in vitro deacetylation assays on acetylated histone tail peptides, analysed by mass spectrometry. b, Western analysis showing SIRT6 deacetylation of H3K9 on full-length histone H3 in vitro. Reactions with NAD+, SIRT6 or the catalytic H133Y SIRT6 mutant protein (SIRT6-HY) are indicated. c, SIRT6 deacetylates H3K9 in cells. Western analysis of 293T cells overexpressing SIRT6, SIRT6-HY or empty vector. d, Western analysis of SIRT6 knockdown in U2OS cells used for T-ChIP in e and f. e, T-ChIP analysis showing hyperacetylation of H3K9 at telomeric chromatin in S6KD U2OS cells. f, Quantification of multiple independent T-ChIP experiments as shown in e. Values represent the T-ChIP signal normalized to input signal, and subtracted for background signal in an IgG control. Error bars indicate s.e.m.; n = 4. P values were determined by two-tailed Student’s t-test.