Fig. 2.

Acquisition of endo H resistance and secretion of SEAP at 2 hours of chase in INS-1 β-cells. Cells were metabolically labeled for 30 minutes and the media (M), cells (C) or total (T, media + cells) were immunoprecipitated with anti-alkaline-phosphatase antibody. At the zero chase time, SEAP had not yet appeared in the medium and all intracellular SEAP was sensitive to digestion with endo H (S). At 2 hours of chase, more than half of labeled SEAP had acquired endo H resistance (R); 35% of labeled SEAP reached the medium (all of which is endo H resistant), and 25% of intracellular SEAP accumulated as an endo-H-resistant form. The data are representative of three experiments.