Immunofluorescence staining analysis of NSC34 and Neuro2a cells overexpressing WT and MT hTDP-43. (A) Physical maps showing the domains of human TDP-43 (hTDP-43) and mouse TDP-43 (mTDP-43). NLS, nuclear localization signal; NES, nuclear export signal; GR, glycine-rich sequence; GQN, glycine (G), glutamine (Q) and/ or asparagine (N)-rich sequence. For hTDP-43, the cDNA inserts in the 3 expression plasmids encoding the WT and the two mutant forms of hTDP-43 are shown. The positions of the nucleotide substitutions are indicated by the stars. (B) Representative photographs showing the immunofluorescence staining patterns of transfected cells. NSC34 (upper two rows) and Neuro2a (lower two rows) cells were transfected with plasmids expressing the WT hTDP-43 and hTDP-43A315T, respectively, and analyzed by immunofluorescence staining at 48 hr post-transfection with use of the 2E2D3 antibody recognizing hTDP-43 and DAPI staining the nucleus. Note the predominant nuclear localization of the exogenous WT and MT hTDP-43 in both types of cells. Scale bar, 10 μm.