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The oocytes were bath-perfused with Ringers solution (115 mM NaCl, 2.5 mM KCl, 1.8 mM CaCl 2 , 10 mM HEPES, and 1 M atropine, pH 7.2) at 4 ml/min for heteromeric nAChR or 2 ml/min for 7 receptors to optimize the measurement of net charge responses (Papke and Papke, 2002)