The mice were anesthetized with sodium pentobarbital. After a laminectomy, the L4-L5 spinal cord was isolated into ice-cold artificial cerebrospinal fluid (ACSF; 119.0 mM NaCl, 2.5 mM KCl, 2.5 mM CaCl2, 1.3 mM MgCl2, 1.0 mM NaH2PO4, 26.0 mM NaHCO3, and 11.0 mM d-glucose, bubbled with 95% O2 + 5% CO2, pH 7.4). The transverse slices (300-μm thickness) were cut with a vibratome. The dorsal quadrants were microdissected out and incubated for 45 min in ice-cold ACSF containing 100 μM Sulfo-NHS-SS-Biotin (Thermo Fisher Scientific, Rockford, IL, USA), followed by brief washes with NH4Cl (50 mM)–containing ACSF. After three washes with normal ACSF, the minislices were lysed in 20 mM tris·HCl (pH 7.5), 50 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% SDS, and 5 mM phenylmethylsulfonyl fluoride (PMSF). After centrifugation at 14,000g for 10 min, the supernatant was collected and incubated with Streptavidin UltraLink Resin (Thermo Fisher Scientific) overnight at 4°C under gentle rotation (25). The resin was washed three times with the lysis buffer and boiled in SDS sample buffer.

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