Matrigel migration chamber

U-2OS cells were cultured in DMEM medium for 48 h. PBS or Chanti-TRIM-treated (80.0 mg/ml) cells were suspended as a density of 1×10⁵ in 500 µl serum-free DMEM for 24 h at 37°C. U-2OS cells were then inserted into the tops of BD BioCoat Matrigel Invasion Chambers (BD Biosciences) according to the manufacturer's instructions. U-2OS cells were incubated with Chanti-TRIM or PBS for 72 h at 37°C using a Matrigel Migration Chamber (BD Biosciences) to analyze the migration of tumors cells. For the invasion assay, a Matrigel Invasion Chamber (BD Biosciences) was used to instead of a Matrigel Migration Chamber. U-2OS tumor cell invasion and migration was measured using a stain-field microscope.

BV-2 (1×10⁶) and BC3H1 (1×10⁶) cells were incubated with tunicamycin (2 mg/ml) for 12 h at 37°C. Cells were suspended at a density of 1×10⁵ in 500 µl serum-free DMEM. For the migration assay, cells were plated in a Matrigel migration chamber (BD Biosciences, Franklin Lakes, NJ, USA) for 24 h at 37°C. For the invasion assay, cells were plated in BioCoat Matrigel invasion chambers (BD Biosciences) for 24 h at 37°C, according to the manufacturer's protocol. Cells were then stained with 5% crystal violet for 30 min at 37°C. Cells were then washed with PBS three times at room temperature and the invasive and migratory tumor cells were counted under a light microscope (Olympus BX51; Olympus Corporation, Tokyo, Japan) in at least three random fields.

Cell migration was detected using the Matrigel migration Chamber (BD Biosciences). GC cells (2 × 10⁵) were seeded into the upper chamber in serum-free DMEM medium. DMEM with 10% FBS was added into the lower chamber. 48 h later, cells that successfully migrated through the chamber membrane were stained with 0.5% crystal violet and then the cell quantity was analyzed.