Qcmtm 24 well fluorometric cell invasion assay kit

Cell invasion was measured using a QCM^TM 24-well Fluorometric Cell Invasion Assay kit (Millipore), which is composed of 85% type I and 15% type III collagen according to the manufacturer’s instruction. Briefly, cells were seeded onto an invasion chamber insert containing an 8-μm pore size polycarbonate membrane coated with a thin layer of polymerized collagen. Invading cells on the bottom of the insert membrane were visualized and counted under the DM 2500 fluorescence microscope.

A rabbit polyclonal antibody for STS was purchased from Abcam (Cambridge, UK). Antibodies against cyclin D1, β-catenin, vimentin, and GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies against GSK3β, phospho-β-catenin (Ser-33/37/Thr-41), and phospho-β-catenin (Ser-45) were purchased from Cell Signaling Technology (Beverly, MA). Anti-phospho-GSK3β (Ser-9), N-cadherin, histone H1antibodies, and QCM^TM 24-well Fluorometric Cell Invasion Assay kit were obtained from Millipore (Bedford, MA). Anti-E-cadherin antibody was from Upstate Biotechnology (Lake Placid, NY). Dual-Luciferase Reporter Assay kit was purchased from Promega (Madison, WI). Ex-Taq DNA polymerase was obtained from TaKaRa Bio (Shiga, Japan). Other chemicals and reagents were of the highest quality commercially available.