Figure 1

SIRT6 interacts with MYH, APE1, and Rad9–Rad1–Hus1. a SIRT6 can be co-immunoprecipitated by hMYH antibody from HeLa extracts. Immunoprecipitation (IP) was performed with hMYH antibody and detected on the Western blot using hSIRT6 or hMYH antibody (lane 2). Lane 1 contains 10% of input cell extracts (IN). Lane 3 is a negative control in which the immunoprecipitation was performed with IgG. b Interaction between hMYH and hSIRT6 is enhanced following H₂O₂ treatment. MYH was co-immunoprecipitated by SIRT6 antibody from extracts prepared from untreated HeLa cells (lanes 4) or from cells treated with 0.15 mM H₂O₂ for 1 h and recovered for 6 h (lane 6). Western blots were detected by hMYH or hSIRT6 antibody. Control lanes are similar to those described in (a). c SIRT6 and APE1 co-immunoprecipitated from HeLa extracts. Immunoprecipitation was performed with hAPE1 antibody and Western blotting was performed with hAPE1 or hSIRT6 antibody. Control lanes are similar to a. d Interaction between hAPE1 and hSIRT6 is enhanced following H₂O₂ treatment. APE1 was co-immunoprecipitated by SIRT6 antibody from extracts prepared from untreated (lanes 4) and H₂O₂ treated (lane 6) HeLa cells. Western blots were detected by hAPE1 or hSIR6 antibody. e, f Immobilized GST-hMYH (~1 μg) and GST-hAPE1 (~0.5 μg), respectively, were used to pull down FLAG-mSirt6 (0.1 μg). Lane 1 contains 10% of input mSirt6 protein. Lane 2 used GST-beads alone. FLAG-mSirt6 was detected by an anti-FLAG antibody. g Immobilized GST, GST-tagged intact hMYH, MYHΔC1, MYHΔC3, MYHΔC3m, and MYHΔC5 (shown in Additional file 1: Figure S2a) were used to precipitate FLAG-mSirt6. The hMYH constructs are depicted.