O-GlcNAcylation performs a critical role in regulating stress response program and cellular homeostasis. However, systematic studies of O-GlcNAc regulated genotoxic stress-responsive transcriptional reprograming have been limited. The GalNAz metabolic labeled global chromatin-associated proteins were isolated from the cross-linked MCF-7 and MCF-7/ADR cells nucleus, and then modified by downstream click-chemistry. The de-crosslinked O-GlcNAz chromatin-bound proteins were enriched for label-free quantitative proteomics by LC-MS/MS.