The described procedure generated data on the initial fluorescence levels of coumarin-30 (i.e., the fluorescence level prior to heating) and normalized MST time traces ("Fnorm" vs. time). For each independent experiment (comprising five ligand conditions, each represented by three technical replicates, all measured on the same capillary tray), the mean initial fluorescence level was calculated for the DMSO control and for each of the four ligand conditions on the same tray.