Fig. 8

SIRT1 co-immunoprecipitates with CDK2 and induces CDK2 deacetylation. a 293 T cells were transfected with pCMV-mCherry, Flag-CDK2, MycTag-SIRT1 for cellular localization analysis (scale = 5 μm). Cells were immunostained with DAPI (blue) for nucleus, CDK2 (green) and SIRT1 (purple). b-c Endogenous immunoprecipitation of SIRT1 and CDK2 was performed in CCRF-CEM cells. d-e Purified GST-tagged SIRT1, CDK2 and HIS-tagged SIRT1, CDK2 were used for in vitro binding assay. f-g Immunoprecipitation was performed using anti-Flag magnetic beads on lysates derived from 293 T cells transfected as indicated. Western blot relative quantification analysis of acetylated CDK2 detected by anti-acetylated-lysine antibody (ac-K). h Immunoblots of SIRT1 and p27 in blood cells from 3 healthy donor and 3 primary T-ALL patient samples with NOTCH1 mutation