Figure 2.

Agonist activation of yeast cells expressing human or rat GPR132. Yeast cells containing a gene‐reporter linked to the pheromone response pathway and engineered to express human GPR132a were treated as follows: Oxidized fatty acid 9‐HODE13 (A); the glycine‐containing ligand, SB‐583831 (C); N‐palmitoylglycine (NPGly; C); N‐linoleoylglycine (NLGly; C); N‐stereoylglycine (NSGly; C); N‐arachidonoylglycine (NAGly; C); N‐oleoylglycine (NOGly; D); linoleamide (D); linoleic acid (D): N‐oleoylserine (NOSer; E) and CP‐55,940 (E). Similarly, yeast expressing rat GPR132 were treated with 9‐HODE (F); N‐palmitoylglycine (NPGly; F); N‐arachidonoylglycine (NAGly; F); SB‐583831 (F); N‐linoleoylglycine (NLGly; G); linoleamide (G) and CP‐55,940 (G). Ligand responses were normalized to the effect of NPGly (for hGPR132a) or 9‐HODE (for rGPR132). Data are presented as mean ± SEM (from n = 3 to 7 independent experiments per ligand; four technical replicates were conducted for each condition). N‐acylamide ligands caused inhibition of yeast cell growth at concentrations >10 µmol/L or >30 µmol/L, and these were therefore used as the top threshold test concentrations curve‐fitting