Figure 8.

Validation of high-throughput quantitative ligand-activated GPCR-induced [Ca²⁺]_i assay using cell monolayers or cell suspensions. (A) Carbachol concentration-dependent activation of M3-AChR-induced [Ca²⁺]_i using HEK293 cell monolayers. Three independent experiments performed with triplicates for each concentration. Baseline [Ca²⁺]_i ranged from 92 to113 nM. α-MSH-concentration-dependent activation of (B) hMC4R or (C) mMC4R (stably expressed in HEK293 cells) -induced [Ca²⁺]_i using cell suspensions. Three independent experiments performed with quadruplicates for each concentration. Baseline [Ca²⁺]_i ranged from 85 to 102 nM. Data shown as mean ± s.e.m.