Nucleosomes reconstituted in the presence of donor chromatin from chicken erythrocytes and fluorescently labeled DNA templates (containing 603 positioning sequence) were gel purified and used for spFRET measurements at a concentration of 0.5 nM, as described (28). Nucleosomes were incubated in the presence of hFACT (0.1 μM) and/or CBL0137 (5 μM) in the TB containing 150 mM KCl for 5 min at 25°C. spFRET measurements and raw data analysis were conducted as described (28).

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