Fig. 3: SIRT6 is a deacetylase of KAT8 and is responsible for KAT8 acetylation.

A HCT116 cells were treated with 1 μM TSA or 5 mM NAM for 8 h and then proteins were extracted for Co-IP assay to detect the level of KAT8 acetylation. B HCT116 cells were co-transfected with myc-KAT8, Flag-SIRT1, Flag-SIRT6 or Flag-SIRT7, and then proteins were extracted for Co-IP to detect the level of KAT8 acetylation. C HCT116 cells were co-transfected with the plasmids above, and then treated with PA at 0.2 mM for 24 h. Proteins were extracted for Co-IP to detect the level of KAT8 acetylation. D, E HCT116 cells were transfected with Flag-SIRT6 (D) or SIRT6 siRNA (E), and proteins were extracted for Co-IP to detect the level of KAT8 acetylation. F myc-KAT8 and Flag-SIRT6 were co-transfected into HCT116 cells, and then KAT8 and SIRT6 proteins were purified to perform in vitro deacetylation assay. G, H HCT116 cells were co-transfected with GFP-SIRT6 (G) or Flag-GCN5 (H) and myc-KAT8, and then treated with PA at 0.2 mM for 24 h. Cell lysates were extracted for Co-IP assay to detect the exogenous interaction between SIRT6 (G) /GCN5 (H) and KAT8. I HCT116 was transfected with GCN5 siRNA or non-specific RNA as control, protein was then extracted for Co-IP to detect the endogenous interaction between KAT8 and SIRT6. J HCT116 SIRT6-KO cells were extracted for Co-IP to detect the endogenous interaction between KAT8 and GCN5.