Figure 10.
Inhibition of UR-CG072 binding to live CHO-K1-hM4R cells by muscarinic receptor ligands. The CHO-K1-hM4R cells (25 000 cells per well) were incubated with 2 nM UR-CG072 and different concentrations of carbachol, arecoline, scopolamine, atropine, pirenzepine, and UNSW-MK259 for 120 min as described in 'Material and methods'. The cell fluorescence intensities were determined by the membrane detection algorithm using the U-Net3-BF-1 model and are presented as individual replicates from a representative experiment performed in duplicate. Four images from different fields of view were measured from a single well. Normalization was done separately for each ligand, 100% corresponds to wells where no competitors were added and 0% corresponds to the wells where the competitors' concentration is the largest.