Western Blot Analysis of WHSC1 and Epigenetic Markers

After treating the OCCC cells with WHSC1-specific siRNAs or GSK126 for the indicated times at the indicated concentrations, total protein was extracted and transferred to a nitrocellulose membrane as previously described [16 (link), 17 ]. Primary antibody diluted with blocking buffer was added to the membrane and reacted overnight at 4 °C. The primary antibodies used in this study were anti-WHSC1 (75,359, Abcam, Cambridge, UK), anti-EZH2 (PA0575, Leica Biosystems, Wetzlar, Germany), anti-H3K36me2 (2901, Cell Signaling Technology, Danvers, MA, USA), anti-H3K27me3 (9733, Cell Signaling Technology), and anti-β-actin (Sigma-Aldrich, St. Louis, MO, USA).