Figure 5.
Effect of PRL-3 mutations on the phosphatase activity and cell migration. A: Catalytic activity of PRL-3 (wild type, D72A mutation, or C104S mutation) proteins in DiFMUP PTPase activity assay. All reactions were performed in triplicate. The data are representative of three independent experiments. B: Quantitative analysis of the cells migrated to the lower side of the membrane. A single-cell suspension (100 μl, 1 × 106 cells/ml) of cells 30 hours after transiently transfection were added into the upper wells of Transwell inserts containing 8-μm-pore polycarbonate membranes precoated with fibronectin on the under surface. Cells were allowed to migrate for 12 hours at 37°C, and then they were fixed and stained. Results represent the average of triplicate samples from three independent experiments. C: Representative expression of PRL-3 mRNA 30 hours after transiently transfection in B16 cells was analyzed by RT-PCR. D: The level of PRL-3 mRNA comparing β-actin mRNA was quantified by densitometry.