Fig. 3
From: O-GlcNAcylation of SIRT1 enhances its deacetylase activity and promotes cytoprotection under stress
SIRT1 is O-GlcNAcylated at Ser549. a ETD-MS detected the O-GlcNAcylated peptide of SIRT1 (544–561). b Either His-tagged wtSIRT1 or SIRT1S549A was coexpressed with or without MBP tagged OGT in E. coli. SIRT1 was immunoprecipitated with anti-His antibody, and then O-GlcNAcylation of SIRT1 was detected by chemoenzymatic labeling and IB with STV-HRP. Data represent three independent experiments. c Flag-tagged wtSIRT1 or SIRT1S549A vector was transfected into NCI-H1299 cells. The cells were lysed after treatment with 2 μM TMG for 4 h, followed by chemoenzymatic labeling and IB. Data represent two independent experiments. d HA-tagged OGT and Flag-tagged wtSIRT1 or SIRT1S549A were co-transfected into NCI-H1299 cells, and the CoIP assays were carried out by anti-Flag antibody, followed by IB with anti-HA and anti-Flag antibodies. Data represent two independent experiments
