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Suppression of synaptic activity was achieved by changing cultured cells into serum-free base-medium (10% minimum essential media (MEM, Life Technologies) and 90% saltglucoseglycine (SGG) containing in mM: 114 NaCl, NaHCO3, 5.292 KCl, 1 MgCl2, 2 CaCl2, 10 HEPES, 1 Glycine, 30 Glucose, 0.5 sodium pyruvate, 0.1% Phenol Red) with 100 nM tetradotoxin (TTX, Tocris) on DIV9 for 24 h to inhibit neuronal action potential firing