To ensure proper synapse formation, primary neurons were kept in culture for 10 days prior to the treatment (Wan
et al,
2012). Then, cells were treated with NMDA (50 μM), NMDA (50 μM) and GI254023x (5 μM), GI254023x (5 μM) alone or vehicle for 30 min. Total cellular mADAM10 levels remained unchanged by the treatment, which is in line with NMDA altering mainly the intracellular localization of ADAM10 by driving the protein to the synaptic membranes (Marcello
et al,
2007). Densitometric quantifications of the Western blots are shown. One‐way ANOVA with
post hoc Dunnett's test (***
P < 0.001,
n = 7). Given are mean ± the standard error of the mean. The mean levels of solvent‐treated cells were set to 1. The membrane was reprobed with the different indicated antibodies.