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In contrast, longer incubation with copper (3 h) produced potentiation of AMPA evoked currents, shifted the concentration-response curve for the agonist (EC 50 = 8.3 μM for control neurons and EC 50 = 2.9 μM for copper treated neurons), and increased the frequency and amplitude of miniature synaptic currents

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Cells were collected at 500 x g for 5 minutes then washed once in PBS