Fig. 1.GluK2 endocytosis is prevented by genistein or
MCD but not by CPZ or pitstop2. Cultured cortical neurons were
pretreated with CPZ (10 mol/L), pitstop2 (25
mol/L), genistein (200 mol/L), or MCD (3
mmol/L) for 30 min before kainate stimulation (300 mol/L, 10 min)
or transferrin binding (20 g/ml, 10 min). (A) The surface and total
GluK2 were immunoblotted with an anti-GluK2/3 antibody. The surface/total ratios
of GluK2 are normalized to that of the control (without kainate stimulation) and
expressed as the mean SD (n = 3). *P 0.05 vs. no kainate.
P 0.05 vs. no inhibitor in kainate stimulation. Neuron
cultures for replicates were made from different animals. (B) The endocytosis
assay of Alexa 488-conjugated transferrin in cortical neurons confirms the
specificity of the inhibitors. Neuron cultures for replicates were made from
different animals.