To purify the human PRC2 core complex, FLAG-tagged EED, 6×His-tagged EZH2, SUZ12, and RBAP48 were cloned into the baculovirus expression vector pFASTBac1 (Invitrogen). Recombinant PRC2 was produced in SF9 cells grown in SF-900 III SFM (Invitrogen). After 60 hours of infection, SF9 cells were resuspended in BC150 [25 mM Hepes-NaOH (pH 7.8), 1 mM EDTA, 150 mM NaCl, 10% glycerol, and 1 mM dithiothreitol (DTT)] with 0.1% NP40 and protease inhibitors [1 mM phenylmethylsulfonyl fluoride (PMSF), 1 mM benzamidine, pepstatin A (1 μg/ml), leupeptin (1 μg/ml), and aprotinin (1 μg/ml)]. Cells were lysed by sonication (Sonic Dismembrator model 100, Fisher Scientific), and PRC2 complexes were purified through Ni-NTA agarose beads (Qiagen), FLAG-M2 agarose beads (Sigma-Aldrich), and Q Sepharose beads (GE Healthcare).

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