Matrigel basement membrane matirx

Cell migration was examined using the transwell chamber (Millipore, Bedford, MA, USA). BD Matrigel™ Basement Membrane Matirx (BD Biosciences, San Jose, CA, USA) was pre-coated to the chamber membrane in the invasion assay, whereas BD Matrigel™ Basement Membrane Matirx (BD Biosciences) was not coated in the migration assay. In brief, transfected U2OS or MG63 cells were harvested and then re-suspended with a serum-free medium. Subsequently, 100 μL re-suspended cells were introduced into the upper chamber, and a relative 500 μL medium with 10% FBS was seeded into the lower chamber. After 24 h of incubation, migratory or invasive cells were analyzed under a microscope (Olympus, Tokyo, Japan).