Inhibition of metastatic properties after transfection with siRNA in cells with expression. (A) Western blotting 72 hours after transfection. The western blotting was also quantified. mock, treatment with Accell siRNA Delivery Media alone; ctr, treatment with siRNA-ctr; si, treatment with siRNA-PRL-3. (B) Anchorage-independent colony formation assays. Representative pictures of colony formation on AZ521 cells were shown in top panel. With mock-treatment as 1.0, the relative rate of colony number was shown in bottom panel. Bars, 200 μm. (C) Proliferation assay. The proliferative activity of AZ521 cells on 1, 2, 3, 4, or 5 days after transfection (top panel) and the relative proliferative rate on 5 days after transfection (bottom panel) were shown. (D) Invasion assay. Cells were seeded on 4 days after transfection, and then incubated for 22 hours. Representative pictures of AZ521 cells (top panel) and the relative invasive rate (bottom panel) were shown. Bars, 200 μm; *, P < 0.05 by ANOVA with post-hoc test; error bars, SD.