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Degradation assay Cells were treated in serum-free medium plus 11 mM glucose agonist (100 nM) before lysis (20 mM Tris base, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 0.5% Triton X-100 plus complete mini EDTA-free protease inhibitor (Roche)), addition of urea sample buffer (100 mM Tris-HCl pH 6.8, 2.5% SDS, 4 M urea, 100 mM DTT, 0.05% bromophenol blue), 10 min incubation at 37 C and immunoblotting