To further investigate this idea that O-GlcNAcylation in mitochondria can improve the efficacy and benefit of mitochondrial transfer, we asked what mitochondrial proteins were modified with O-GlcNAc. To identify O-GlcNAcylated proteins and the potential target sites of O-GlcNAcylation, rat astrocytes were stimulated with NAD to induce O-GlcNAcylation in mitochondria, then we isolated mitochondria and extracted O-GlcNAcylated proteins by immunoprecipitation using O-GlcNAc antibody followed by silver staining.