Fig 3. NRP2 downregulation in FB-2 cells inhibits cell proliferation, migration, and invasion.

(A) NRP2 downregulation in PAX8 clones was analyzed by Western blot with a specific anti-NRP2 antibody. (B) Growth curves of FB-2, FB2-FLAG, FB2-P8cl25 and FB2-P8cl31 cells are shown. Triplicate of 8 x 10⁴ cells were seeded into 60-mm plate. Cell numbers were counted on days 1, 2, 3 and 4 after seeding. (C) Wound-healing migration assay for FB-2, FB2-FLAG, FB2-P8cl25 and FB2-P8cl31 cells were performed. The healing of the wounds by migrating cells was imaged at time 0, 8 and 16 h. (D) Matrigel invasion assay of FB-2, FB2-FLAG, FB2-P8cl25 and FB2-P8cl31 cells. The cells were seeded on 8 μm pore size Transwell filters and allowed to migrate toward 10% fetal bovine serum. After 16 h, the upper surface of the filter was wiped clean and cells on the lower surface were stained and photographed. This figure is representative of three independent experiments.