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For NMJ analysis, 20 m thick abductor digiti minimi muscle cryosections were blocked with 5% bovine serum albumin (Sigma-Aldrich, St Louis, MO, USA)/0.1% Triton X-100 in PBS then immunostained with with CF488 conjugated--bungarotoxin (00005, Biotium, Fremont, CA, USA) for visualizing nicotinic acetylcholine receptor (AChR) of neuromuscular junctions and anti-NF-200 (1:2000, N4142, Millipore Sigma) for visualizing axons for 1 h at RT or overnight at 4 C