In vitro heat-denatured luciferase refolding assay

Luciferase refolding assay was performed as previously described [28,29] with minor modifications. Briefly, 80 nM luciferase, 160 nM DnaJ, and 400 nM Hsp70 (1:2:5) in buffer (25 mM HEPES/pH 7.4, 50 mM KCl, 3 mM MgCl₂, 2 mM DTT) with 2 mM Mg-ATP were heat-denatured at 43°C for 10 min in the presence or absence of AptHsp70-1 (Hsp70:Aptamer = 1:4). The mixture was incubated at room temperature for various time periods ranging from 1 to 150 min. Luciferase activity was measured as luminescence using luciferase assay reagent (Promega) according to the manufacturer's instructions.