FIGURE 1.
Effects of serotonin on 5-HT2AR trafficking. A, serum-deprived HEK293 cells, transiently transfected with 5-HT2AR-v5 for 24 h, were incubated on ice with 1 μm 5-HT for 60 min, washed free of unbound ligand, and exposed to prewarmed ligand-free medium at 37 °C for 0, 10, 30, and 180 min. Cells then were fixed, stained with anti-v5 antibody (visualized with Alexa Fluor 568-conjugated secondary antibody (red)), and analyzed by confocal microscopy. The bar is 5 μm. B, serum-deprived HEK293 cells, which either were NT or transiently transfected with 5-HT2AR-v5 for 24 h, and treated at 37 °C with 1 μm 5-HT for 0 or 180 min. After being washed with ice-cold PBS, cell lysates were subjected to immunoprecipitation (IP) with an antibody to 5-HT2AR-v5, followed by immunoblotting (IB) with an antibody to 5-HT2AR-v5. Data shown in panels A are representative of three independent experiments. Results in panel B are mean ± S.E. of three separate experiments.