Cell differentiation assay

N2a, IMR32 and SHSHY-5Y cells were seeded at a density of 5000 cells/well into a 24-well culture plate in complete culture medium (400μL/well) and incubated under standard culture conditions for 24h. Complete culture medium was aspirated and wells were washed with PBS before treatment with serum-free culture medium (SFM) containing FPRa14 (0–10μM). Cells treated with SFM alone served as a negative control. The cells were then incubated under standard culture conditions for 48h in order to observe any morphological changes. For cell differentiation assays with FPR antagonists, cells were pre- incubated with Boc-MLF (0–40μM), cyclosporin H (0–40μM) or WRW4 (0–40μM) for 30min before addition of FPRa14 (8μM).