CUT&RUN Sequencing of EZH2 binding sites in HEK293T cells expressing a Dual-specificity Aptamer that specifically increases O-GlcNAcylation on beta-catenin when O-GlcNAc transferase was inhibited

O-GlcNAc is a dynamic post-translational modification on thousands of intracellular proteins, it regulates protein functions and is involved in many metabolic diseases. Using a dual-specificity aptamer, we targeted the O-GlcNAc transferase (OGT) to endogenous ß-catenin and specifically increased O-GlcNAcylation of ß-catenin. We found that O-GlcNAc on ß-catenin promoted its interaction with EZH2 regardless of the Wnt signaling status. To study how this interaction regulates the distribution of EZH2 on the genome, we performed Cleavage Under Targets and Release Using Nuclease (CUT&RUN) on HEK293T cells expressing individual (Ctrl.) or dual-specificity aptamers, under Wnt - and Wnt + conditions. This is a control experiment in which all samples were treated with the OGT inhibitor Ac5S. Overall design: 12 samples from 4 groups of HEK293T cells were analyzed: cells expressing Ctrl (T1 · bc339) or Dual-specificity (3JB8F+12) aptamers were exposed to Wnt- or Wnt+ medium. Each group contains 3 replicates. Cells in all replicates were treated with an OGT inhibitor (Ac5S).