JZL195 was prepared as saline-emulphor emulsions by vortexing, sonicating, and gentle heating neat compound directly into an 18:1:1 vol/vol/vol solution of saline:ethanol:emulphor. JZL195 was administered as milky suspensions. Male C57BL/6J mice (< 6 months old, 20–28 g) were i.p. administered compound or an 18:1:1 vol/vol/vol saline: emulphor:ethanol vehicle at a volume of 10 mkl/g weight

CB1 receptor transfected CHO cell membranes were thawed on ice, centrifuged at 50,000 g for 10 min at 4 °C, and resuspended in Assay Buffer A (50mMTris-HCl (pH 7.4), 3mMMgCl2, 0.2mM EGTA, and 100 mM NaCl). Reactions containing 10 mkg of membrane protein were incubated for 90 min at 30 °C in Assay Buffer A containing 10 mkM GDP, 0.1 nM [35S]GTPgammaS, 0.1% BSA, and various concentrations of JZL 195.