Fig. 2.

Blocking GPR110 activation abolishes the anti-inhibitory effects of synaptamide in microglia. Cells were incubated with LPS (100 ng/mL) for the indicated time, and GPR110 mRNA levels were determined by qPCR, showing LPS-induced elevation of GPR110 in BV2 murine microglia cell line, 8-week-old mice-derived primary adult microglia and 8-week-old mice whole brain (a). Control IgG and GPR110 N-terminal blocking antibody (300 ng/mL) were added 30 min prior to LPS treatment (100 ng/mL). Synaptamide (10 nM) was added immediately after LPS exposure. The mRNA levels of TNF and IL-1β were determined by qPCR after incubation for 1 h (b). After 24 h, accumulated levels of TNF and IL-1β in the media were measured by ELISA (c). Cells were treated with 10 nM synaptamide or 10 μM forskolin (Fors) as a positive control for 15 min, and cAMP levels were measured. Translocation of RelA (p65) was determined in BV2 cells by immunocytochemistry (red, RelA; blue, DAPI) (e). The data are expressed as the mean ± SEM (n = 3) representing at least three independent experiments. ns, the difference of means is not statistically significant