(A) Double-immunolabeling of spinal cordneurons with ChAT (red) and N-terminal FUS antibody (green). Nuclei were visualized with DRAQ5 (blue). Cellular localization of FUS was analyzed in the ventral spinal cord of Fus+/+/ChAT-CRE (A1-A4), FusΔNLS/ΔNLS/- (A5-A8), and FusΔNLS/ΔNLS/ChAT-CRE (A9-A12).FUS was completely nuclear in ChAT+ neurons of Fus+/+/ChAT-CRE, while cytoplasmic in FusΔNLS/ΔNLS/-, In the ventral horn of FusΔNLS/ΔNLS/ChAT-CREmice, ChAT+ neurons (motor neurons, e.g. within the dashed square) displayed nuclearFUS immunoreactivity, while ChAT negative cells retained cytoplasmicFUS immunoreactivity (arrows) 