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Compound 25 as an inverse agonist of hCB1. CHO-hCB1 cells were loaded with a calcium indicator dye for 60 min as described in the Experimental Section. The cells were then stimulated by various concentrations of each compound, and the fluorescence changes were recorded using a FLIPR Tetra (Molecular Devices) plate reader for 60 s. Cells stimulated with only the vehicle in the buffer served as a negative control. Compound 1 served as a positive control for this experiment.
