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ATPase assays were performed utilizing an ATP-regenerating system51. Assays were measured at 30 °C in 40 mM HEPES, 150 mM KCl, 5 mM MgCl2, pH 7.5 (standard buffer) and 2 mM ATP, supplemented with 50 µM DEX using a Varian Cary50 Bio UV-Vis spectrophotometer (Varian Inc., Palo Alto, USA). The GR-LBD was pre-incubated with 3 µM yeast wt or mutant Hsp90 for 10 min. Background ATPase activity was determined by the addition of 50 µM of the Hsp90 inhibitor Radicicol.
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