Updated project metadata. To find the SIRT6 and SIRT7 binding proteins, we expressed V5-tagged SIRT6 and SIRT7 and pulled down this protein with anti-V5 agarose affinity gel. Sirtuins are a family of NAD-dependent deacetylases which deacetylate not only histones but also a wide range of target proteins. Sirtuins are conserved from yeast to mammal, and are known to regulate many processes such as cellular metabolism, apoptosis, cellular senescence, cell cycle, and organism-level aging. Among the members of sirtuins showing different subcellular localization, SIRT1, SIRT6 and SIRT7 are localized in nucleus. Whereas many functions and interacting proteins of SIRT1 have been studied, functions of SIRT6 and SIRT7 are not extensively revealed yet. Since it is important to understand the molecular function of SIRT6 and SIRT7, we here aimed to identify interacting proteins in mammalian cells. Affinity purification and nano-liquid chromatography/tandem mass spectrometry were used for exploring novel SIRT6 and SIRT7-interacting partners which were validated by co-immunoprecipitation, immunoblot and confocal microscopy. Taken together, our study has identified novel SIRT6 and SIRT7-interacting proteins. Bioinformatics pipeline: We performed peak list generation of MS data, and identification/quantification of peptides and proteins from three technical replicates of LC-MS/MS data using MaxQuant quantification tool with Andromeda search engine (version 1.3.0.5) (Cox and Mann, 2008; Cox et al., 2009; Cox et al., 2011). Top 10 peaks per 100 Da were used for analysis. Enzyme specificity for trypsin was used. The minimal peptide length was six amino-acids, and we allowed two mis-cleavages. Variable modification options were used for the oxidation of methionine (15.995 Da), and carbamidomethylation of cysteine (57.021 Da). The tolerance was set to 10 ppm for precursor ions and 0.8 Da for fragment ions. SwissProt database (Homo sapiens reference proteome set, release 2013_01, 20,226 entries), to which contaminants and reverse sequences were added. For peptide and protein identification the one percent false discovery rate (FDR) was determined by accumulating 1% of reverse database hits. In the case of identified peptides that are all shared between two proteins, these are combined and reported as one protein group. For comparison between samples we used label-free quantification with a minimum of two ratio counts to determine the normalized protein label-free quantification (LFQ) intensity (Luber et al., 2010).