ThT fluorescence from the aggregation mixture was monitored on a plate reader (Analyst AD; Molecular Devices, Sunnyvale, CA) using an excitation wavelength of 440 nm and an emission wavelength of 480 nm as previously reported [21 (link)]. Two aliquots (~260 μL technical replicates) from the aggregation reaction (containing 20 μM ThT) were placed on a black 96-well polystyrene plate (Greiner, Monroe, NC) at specific time points to measure the fluorescence intensity. After the measurements, aliquots were placed back into the microcentrifuge tube to resume the incubation at 37°C with orbital shaking set at 300 RPM. The measurement process was repeated until the apparent change in fluorescence intensity plateaus over a period of at least 72 h.