Fig 1. The schematic depiction of the peptide microarray process for discovering OGT substrates.

The microarray was blocked with BSA, and followed by the addition of OGT (s-OGT, m-OGT, and nc-OGT), UDP-GlcNAc, and a FITC conjugated antibody. The enzymatic reaction was run by pumping the mixture up and down through the porous Al₂O₃ chip material. Images of the fluorescent signals were generated every 10 minutes during 4 hours (kinetic readout). Images were quantified using Bionavigator 6 software.