Fig. 2.
The effects of the PAC1 receptor antagonist PACAP6-38 on SH-SY5Y cells. SH-SY5Y cells were maintained in low serum medium as described in Materials and methods. (a) Histogram showing fold change in cAMP levels for cells treated for 15 min with 100 nmol/L PACAP-38, 1 μM VIP or 1 μM Ro 25-1553, in the presence (dark gray bars), or absence (light gray bars) of 10 μM PACAP6-38. The basal level was determined for cells maintained in low serum medium alone. (b) Histogram showing numbers of neurite-bearing cells (fold of basal control) following treatment for 4 days with 100 nmol/L PACAP-38, 1 μM VIP or 1 μM Ro 25-1553, in the presence (dark gray bars) or absence (light gray bars) of 10 μM PACAP6-38. Standard error bars are shown (n = 3) and one-way ANOVA performed to identify significant differences between treatments (***p < 0.001, **p < 0.01, *p < 0.05). (c) Western blot analysis of cell extracts using antibodies specific for Bcl-2 and GAP-43 was carried out following the different treatments for 4 days. Protein loading was determined using an antibody specific for GAPDH. Each blot represents three independent experiments. (C = control, p = 100 nmol/L PACAP-38, V = 1 μM VIP, R = 1 μmol/L Ro 25-1553).