Figure 3

Pharmacological blocker of the glycine binding site prevents glycine primed NMDAR endocytosis. A, Top Representative Western blot of co-immunoprecipitation of NMDARs with adaptin β2 from HEK293 cells expressing recombinant wild type NMDARs treated with ECS, glycine (100 μM) with or without L689560 (1μM), or with L689560 alone. Bottom, Histograms showing quantification of GluN1 and adaptin β2 association after glycine stimulation with or without L689560 (n = 4). Data are normalized to ECS group and presented as mean percent of ECS control (± s.e.m). B, NMDAR internalization measured by cell ELISA assay. Cultures (n = 5) were pre-treated with glycine (100 μM) plus APV (100 μM) with or without L689560 (1μM) followed by NMDA (50 μM) plus glycine (1μM). * indicates p <0.05, ** indicates p < 0.01 compared to L689560-treated group.