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Intracellular COMP is an Endo H-sensitive glycoprotein. COS7 cells were transfected with wild-type or mutated COMP and cultured at 37°C. The cells were saved at 24 hours and 48 hours and solubilized with 0.5%SDS/1% 2-mercaptoethanol/50 mmol/L sodium acetate (pH 5.5), containing protease inhibitors after washing with PBS. The samples were incubated in the presence or absence of Endo H for 3 hours. Treated samples were analyzed by immunoblotting with the anti-COMP antibody.
