Biosynthetic pathway of insulin-HT monitored by fluorescent microscopy.

<p>MIN6 cells transiently transfected with insulin-HT plus calnexin-YFP, insulin-HT plus M6PR-EGFP, or insulin-HT plus phogrin-EGFP were treated with blocking probe for 1 hour. After removal of excess probe, the cells were labeled with HT-TMR for 1 hour at 15°C (top panels). The cells were subsequently chased at 37°C for 15 (middle panels) and 30 minutes (bottom panels), respectively. Fluorescent images were analyzed by confocal microscopy. Experiments were repeated three times with reproducible results. Bar, 10 µm.