Figure 2.

C10 and other ligands can activate GPR84. Flp-In T-REx 293 cells harboring FLAG-hGPR84-eYFP at the Flp-In T-REx locus were untreated (− dox) or maintained in the presence of doxycycline (100 ng/ml) for 24 hours. Such cells were imaged to detect the eYFP tag linked to the receptor (a). The capacity of fatty acids of chain length 9–14 (100 μM) to inhibit forskolin-stimulated cAMP levels in doxycycline-induced cells was assessed and compared to effects of embelin and DIM (b). C10, embelin, DIM and compound 1 each regulate cAMP levels in a concentration-dependent manner (c). Membrane preparations generated from doxycycline-induced cells were also used in [³⁵S]GTPγS binding assays and each of C10, embelin, DIM and compound-1 promoted binding of [³⁵S]GTPγS in a concentration-dependent manner (d). By contrast methyl decanoate was without effect (d).