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Pulse-chase labeling of CHO cells that express wild-type and mutant insulin. CHO-Ins2wt (clone w9, upper) and CHO-Ins2Mody (clone a7, lower) were labeled for 30 min with [35S]methionine. After chasing for indicated times, immunoprecipitation was performed from either cell extracts (C) or media (M), using a mixture of anti-insulin and anti–C-peptide antibodies. Immune complexes were analyzed with tricine–SDS-PAGE (16.5% polyacrylamide gel).
