Figure 4.
The N-termini and C-termini of Plg-RKT are exposed on the cell surface. Membrane fractions of PC12 cells stably transfected with V5-pCIneo-Plg-RKT were incubated with either buffer (lane 1), trypsin (1 mg/mL) (lane 2), or trypsin 1 mg/mL + soybean trypsin inhibitor (SBTI) (2 mg/mL) (lane 3) for 30 minutes at 37°C or intact PC12 cells were incubated with 1 mg/mL trypsin for 2 hr at 37°C, followed by 2 mg/mL SBTI for 15 min. Following neutralization of trypsin with SBTI, the membrane fraction was prepared from the treated, intact cells (lane 4). 30 μg/lane of membrane fractions was electrophoresed on 18% SDS page under reducing conditions and western blotted with either anti-V5, anti-Plg-RKT mAb, or isotype control. This research was originally published in [17]