Figure 1
Chemical structures of AEA and NAGly. The chemical structures of N-arachidonoyl glycine (NAGly) and its precursor, the endocannabinoid N-arachidonoyl ethanolamide (Anandamide; AEA). Enzymatic pathways for endogenous AEA – to NAGly conversion were previously identified [13].
Figure 2
Cell migration induced by NAGly, O-1602, Abn-CBD. Cell migration in response to Vh (0.1% DMSO); 1 μM NAGly; 1 μM O-1602; 1 μM Abn-CBD in BV-2, HEK293-GPR18 and HEK293 WT cells. n = 3. This figure represents data previously published [11] and is intended to illustrate the context of the novel findings reported here.
Figure 3
GPR18 mRNA levels in siRNA transfected BV-2 microglia. Gel electrophoresis of HEK293-GPR18, HEK293 WT, Normal BV-2 microglia and GFP+ GPR18 siRNA transfected BV-2 microglia RT-PCR products run on a 2% agarose gel.
Figure 4
Immunocytochemical staining of GPR18 receptors in siRNA transfected BV-2 microglia. Immunofluorescent confocal microscopy was conducted using an antibody against the GPR18 C-terminus (1:150; green), phalloidin to label actin (1:40; red). Normal BV-2 microglia with: a) phalloidin b) GPR18 antibody c) phalloidin and GPR18 antibody. GFP+ GPR18 siRNA transfected BV-2 microglia with: d) phalloidin e) GPR18 antibody f) phalloidin and GPR18 antibody.
Figure 5
Cell migration induced by NAGly, O-1602, Abn-CBD in wildtype BV-2 microglia (lefthand bars) and siRNA transfected BV-2 microglia (righthand bars). Cell migration in response to Vh (0.1% DMSO); 1 μM fMLP; 1 μM NAGly; 1 μM O-1602; 1 μM Abn-CBD in Normal BV-2 microglia and GFP+ GPR18 siRNA transfected BV-2 microglia. n = 3; *** = P < 0.001, one-way ANOVA.