Transwell migration and invasion assays were carried out in 24-well modified Transwell Boyden chambers (BD Transduction, Franklin Lakes, NJ, USA). The upper surface of 6.4-mm-diameter filters with 8-µm pores was precoated with (invasion assay) or without (migration assay) Matrigel (BD Transduction). The siRNA transfectants (8 × 105 cells per well) were seeded into the upper chamber with serum-free medium. Complete growth-medium was added to the lower well of each chamber. The transfectants were incubated for 22 h and then migrated or invasive cells on the lower surface of the filters were fixed and stained with Diff-Quik stain (Sysmex, Kobe, Japan). The stained cell nuclei were counted directly in triplicate, as described elsewhere 37-39.

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