Transwell cell migration assay kits

Cell viability was determined using an XTT assay. Briefly, approximately 1,000 - 2,000 cells were subcultured in a 96-well plate. XTT was measured at days 0, 1, 2 3 and 4 post-plating using the Cell Proliferation Kit II (Roche Biosciences, Indianapolis, IN, USA). Cell migration and invasion assays were performed using Transwell cell migration assay kits (Corning, New York, NY, USA) and BioCoat™ Matrigel® Invasion Chambers (Corning), respectively. For doxycycline-induced miR-193a-3p expression, cells were pretreated with or without 1 μg/ml doxycycline. After 48 h, the cells were trypsinized and placed in the upper chamber filled with serum-free culture medium with or without doxycycline (1 μg/ml) and allowed to migrate or invade.

Cell viability was investigated using Cell Counting Kit-8 and colony formation assays. Cell migration and invasion assays were performed using Transwell cell migration assay kits and BioCoat™ Matrigel^® Invasion Chambers (Corning Incorporated), respectively. For the migration assay, 5×10⁴ cells were seeded into the upper chamber and incubated for 24 h at 37°C in 5% CO₂. For the invasion assay, 1×10⁵ cells were plated on chambers pre-coated with 1.6 mg Matrigel (Corning, Inc.) and incubated for 36 h at 37°C in 5% CO₂. Subsequently, cells were fixed, stained with 0.5% crystal violet solution at room temperature for 10 min, photographed and counted under a light microscope (magnification, ×200).
Cell apoptosis was analyzed with the Annexin V-fluorescein isothiocyanate/propidium iodide Apoptosis Detection kit. Briefly, 1×10⁵ cells were re-suspended in 500 µl binding buffer and stained with 5 µl Annexin V and 5 µl PI in the dark at room temperature for 20 min. The samples were then analyzed using a FACScan flow cytometer and the CellQuest™ Pro 1.0 software (BD Biosciences), according to the manufacturer's instructions.