Western Blot Analysis

Whole cell lysates, nuclear and cytoplasmic extracts, or eluted fractions from co-IP experiments were separated on a 4–12% Bis-Tris gradient gel (Invitrogen) followed by transfer onto a 0.2 µm polyvinylidene difluoride membrane. Primary antibodies used for western blot analysis were targeted against H3K27me3, histone H3, FLAG-tag, EZH2, SUZ12, embryonic ectoderm development (EED), β-tubulin, lamin B1, or β-actin. Secondary antibodies used were goat anti-mouse horseradish peroxidase (HRP) and goat anti-rabbit HRP. Chemiluminescence was detected using the Chemostar ECL Imager device (Intas Science Imaging). A detailed protocol regarding sample preparation and antibodies utilized can be found in the Supplementary Material.