For microglial migration evaluation, Transwell assays were performed using 8-μm-pore-diameter inserts (MCEP24H48, Millipore, Darmstadt, Germany). Briefly, 2 × 105 primary microglial cells were plated in the upper chamber with 200 μl of serum-free medium, and the chamber was then placed within the bottom wells (24-well plate) containing 600 μl of normal medium supplemented with or without NaR (20 g/ml) or 5 μM Aβ as indicated, followed by incubation for 24 hours. The nonmigrating cells on the upper membrane were removed with a cotton swab, and the cells on the lower surface of the membrane were fixed with 4% paraformaldehyde for 15 to 30 min. After staining with 0.2% crystal violet (30 min at room temperature), images were randomly captured, and the number of migrating cells were manually quantified.

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