Research Paper Volume 1, Issue 1 pp 109—121

Figure 1. SIRT6 interacts with the DNA-PK DSB repair factor. (A) Endogenous SIRT6 protein associates with large molecular weight complexes. HeLa cell nuclear extract (NE) was separated by gel-filtration and fractions subjected to Western analysis with SIRT6 antibody. Fractions with molecular weight standards are indicated (arrows). (B) Coomassie stain of proteins in Flag-SIRT6 or negative control IPs from 293T cells. The identities of proteins detected by mass spectrometry are indicated. (C) SIRT6, but not SIRT1, associates with DNA-PK subunits. Western analysis of SIRT6, SIRT1, and negative control IPs with the indicated antibodies. WCE: whole cell extract. (D) SIRT6 interaction with DNA-PKcs, but not Ku70 and Ku80, is resistant to ethidium bromide (EtBr). Western analysis as in (C) except EtBr was added as indicated to disrupt DNA-mediated interactions. (E) Endogenous interaction between SIRT6 and DNA-PKcs. Western analysis of SIRT6-bound proteins in co-IPs from 293T cells.