FIGURE 4.

Metabolic labeling approach identifies O-GlcNAc proteins from human T cells. (A) Diagram illustrating the workflow for identification of O-GlcNAc proteins from T cells using metabolic labeling. (B) Primary human T cells from one donor were metabolically labeled with GalNAz and stimulated with control beads or anti-CD3/CD28 beads for 24 h. Azide-labeled proteins were then biotinylated for subsequent enrichment by streptavidin beads. Samples of the input (In) and flow-through (FT) material were analyzed by immunoblot to detect the biotin tag. Biotinylated protein was eluted (E) from an aliquot of the streptavidin beads to confirm capture.