Anti histone h1

Total protein was extracted from the collected cells. Cytoplasmic and nuclear proteins were extracted using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, Rockford, IL, USA) according to the manufacturer’s instructions. Equal amounts of protein were separated via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and were then transferred onto polyvinylidene fluoride (PVDF) membranes (Merck Millipore). Membranes were subsequently blocked for 2 h in Tris-buffered saline containing 0.1% Tween and 5% nonfat dry milk followed by incubation with primary antibodies overnight at 4°C. After a second incubation with horseradish peroxidase-conjugated secondary antibodies (1:5000; Multi Sciences Biotech, Hangzhou, China), the bands on the PVDF membranes were developed using enhanced chemiluminescence (ECL; Biological Industries, Beit HaEmek, Israel). All primary antibodies (anti-γ-catenin, anti-β-catenin, anti-Cyclin D1, anti-c-Myc, anti-GSK3β, anti-phospho-GSK3β, anti-Histone H1 and anti-β-actin) were obtained from Cell Signaling Technology (Beverly, MA, USA).