BL21(DE3) plysS–competent cells bearing plasmids encoding GST, GST–N-Plk1, or GST–MMSET-SET were induced with 0.2 mM isopropyl-β-d-thiogalactopyranoside (IPTG) at 30°C for 4 hours. BL21(DE3) pTf16–competent cells bearing a plasmid encoding GST–G9a-SET was induced with 0.1 mM IPTG and l-arabinose (0.5 mg ml−1) at 16°C for 8 hours.

For protein expression and purification in Sf9 insect cells, G9a-SET construct was subcloned into a pFastBac-GST plasmid. Recombinant baculoviruses were generated according to the manufacturer’s protocol (Invitrogen). When expressing proteins at large scale, the Sf9 cells were grown in an orbital shaker at 27°C at a constant stirring rate of 100 rpm in the dark. The baculoviruses were added to the Sf9 cells at a density of 1 × 106 cells ml−1, and the cells were harvested around 72 hours after transduction. Cells were lysed in buffer containing 50 mM tris-HCl, 150 mM NaCl, and 0.05% NP-40, and proteins were purified using a glutathione-Sepharose resin (Aogma), followed by elution with glutathione [20 mM in 75 mM tris-HCl (pH 8.0)] and dialysis.

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