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Immunofluorescence staining of cultured neurons To determine the effect of lithium on neuronal morphology, neuron cultures were fixed with 4% paraformaldehyde for 30 min at room temperature, permeabilized with 0.25% Triton X-100 in phosphate buffer saline for 10 min, blocked with 5% donkey serum (Jackson Laboratories, PA) for 1 h at room temperature, and then incubated for 16 h at 4C with rabbit anti-microtubule-associated protein (MAP)2 antibody (Millipore, MA)

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Initially, the 2,7-dichlorofluorescein diacetate (DCFH-DA) probe was used to detect intracellular ROS generation