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Rapid dilution assay to test reversibility of C34 interaction with cathepsins. Recovery of enzymatic activity was measured after incubation of each enzyme in the absence (Control) or presence of inhibitor (C34). Initial rates were measured for 10–20 min after dilution of the enzyme-inhibitor complex with substrate to initiate reaction, and plotted as Relative Fluorescence Units (RFU) of AMC released from substrate per unit of time (s).
