1 mg of nuclear extract was diluted 1:4.2 with buffer C-0 (20 mM Hepes/KOH pH 7.6, 1.5 mM MgCl2, 0.2 mM EDTA, 25% (v/v) glycerol, 0.131% (w/v) NP-40, 1 mM DTT) and adjusted to 1 ml final volume with buffer C-100 (20 mM Hepes/KOH pH 7.6, 100 mM NaCl, 1.5 mM MgCl2, 0.2 mM EDTA, 25% (v/v) glycerol, 0.1% (w/v) NP-40, 1 mM DTT). 5 U/ml of Benzonase was added (70664, Millipore), samples were incubated for 1 h at 4°C with rotation and diluted extracts were cleared of contingent precipitates by centrifugation (15 min, 21,100 g, 4°C). 25 μl of GFP-Trap Agarose (gta, ChromoTek) or ANTI-FLAG M2 Affinity Gel (A2220, Sigma) was blocked in buffer C-100 containing 1 mg/ml BSA and 1% (w/v) fish skin gelatin for 1 h at 4°C with rotation and then added to the diluted extracts. Immunoprecipitation was carried out overnight at 4°C with rotation. The resin was washed four times with 1 ml IP150 buffer (25 mM Hepes/KOH pH 7.6, 150 mM NaCl, 12.5 mM MgCl2, 0.1 mM EDTA, 10% (v/v) glycerol 0.1% (w/v) NP-40, 1 mM DTT) and finally resuspended in SDS-PAGE loading buffer (50 mM Tris/HCl pH 6.8, 2% (w/v) SDS, 10% (v/v) glycerol, 0.1% (w/v) bromophenol blue, 100 mM DTT). Immunoprecipitates were analysed by SDS-PAGE and Western blot.

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