FIGURE 7.

Labeling of cell surface-exposed GPR17 with an antibody raised against the N terminus of GPR17. Cells were cultured with CM for 48 h and then incubated for 45 min at 4 °C with the anti-Nt-GPR17 antibody without or with preincubation with the antigen. After washing, the cells were fixed and labeled with a monoclonal antibody against MAG followed by an incubation with anti-mouse IgG conjugated to Cy3 (MAG, red) and an anti-rabbit IgG conjugated to fluorescein to reveal the distribution of the anti-GPR17 antibody (GPR17, green). Note that anti-Nt-GPR17 labels the surface of cells that do not express or express small amounts of MAG. Immunostaining for GPR17 was completely abolished by preincubating the antiserum with the peptide used for rabbit immunization. The merged images are shown. Bars = 10 μm.