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Overall protein O-GlcNAcylation in cytosolic and mitochondrial fractions. Cytosolic and mitochondrial proteins from mouse hearts were separated and analyzed by Western blot. 25 μg of cytosolic or 40 μg of mitochondrial proteins for each lane were loaded. The membrane was probed with anti-O-GlcNAc antibody (RL2), stripped and reblotted with anti-SERCA or an antibody for respiratory chain complex IV subunit I (anti-COX I).
