CTHRC1 and β-Catenin Expression Analysis

Western blotting was performed as previously described [12 (link)]. The antibodies for CTHRC1 (Abcam, Ltd., USA), β-catenin, GAPDH and LaminB1 (Abcam, Cambridge, UK) in 5% milk/TBST (tris-buffered saline Tween-20) were used. The signal was observed by using an enhanced chemiluminescence (ECL) plus kit (Milipore). The relative expression of each protein was determined using NIH Image J software. As for the immunofluorescence assay, cells cultured in the chamber slides were probed with CTHRC1 and β-catenin according to the manufacturer's instructions. The florescein isothiocyanate (FITC)-coupled and Rhodamine (Rhod)-coupled secondary antibodies were purchased from Molecular Probes. Treated cells were observed and pictured through an Olympus BX51 fluorescence microscope (Olympus, Tokyo, Japan).