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Figure 5


Fig. 5. Endogenous expression of Cab45, a Golgi lumenal-resident protein, in INS-1 cells. Cells were pulse labeled for 30 minutes with 35S amino acids and chased continuously for 4 hours in the absence (-) or presence (+) of secretagogue (Stim). In duplicate, media (Med.) were collected and cells lysed, and the samples were then analyzed by sequential immunoprecipitation with mAb anti-Cab45 (lower panel) and polyclonal anti-insulin (upper panel). The Cab45 immunoprecipitates were further divided either for mock digestion (-) or endoglycosidase H (Endo H) digestion (+). At 4 hours of chase, Cab45 appears as two intracellular endo-H-resistant bands; the position of the 45 kDa molecular mass marker is shown.





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