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Subsequently, the brains were collected and postfixed in paraformaldehyde for 48 h at 4 C and sectioned to obtain 12-m slices, which were permeabilized with 0.05% Triton X-100 (Vetec, Speyer am Rhein, Germany) solution in PBS for 30 min and subsequently incubated with blocking solution containing 5% bovine serum albumin (SigmaAldrich), 2.5% normal goat serum (Thermo Fisher Scientific, Waltham, MA), and 0.02% Triton X-100 diluted in PBS for 1 h
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If the break exceeds four weeks, most protocols recommend restarting the full titration from 0.25 mg
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