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REAGENTS/MATERIAL:
Antibodies used in this study include:
rabbit anti-C-terminal NR2B, (Sigma Aldrich, St. Louis, MO),
anti-synaptophysin, (Sigma Aldrich) and
mouse monoclonal anti-CK2, subunit β (Sigma Aldrich).
rabbit phosphorylation state-specific Serine1480 anti-NR2B (Pierce Thermo Scientific, Rockford, IL).
rabbit anti-NR2A (Upstate, Lake Placid, NY).
anti-actin, (Applied Biological Materials, Richmond, BC, Canada), (used for immunoblotting loading control).
mouse monoclonal anti-EEA1, (BD Biosciences, San Jose, CA).
anti-GluR1 (Chemicon, Billerica, MA) and
rabbit phosphorylation state-specific Tyrosine1472 anti-NR2B (Chemicon).
anti-NR1, (Affinity BioReagents, Golden, CO),
mouse monoclonal anti-PSD-95, (Affinity BioReagents) and
mouse monoclonal anti-CK2 subunit α, (Affinity BioReagents).
Anti-GFP and all secondary antibodies for immunofluorescence were obtained from Invitrogen (Molecular Probes, Eugene, OR).
All the drugs and inhibitors used in this study were purchased from Tocris Cookson (Ellisville, MO) except APV and picrotoxin (Sigma) and DRB (Calbiochem, San Diego, CA).
GFP-NR2A and GFP-NR2B constructs were kindly provided by Dr. Stefano Vicini (Georgetown University).
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