Overexpression of RANBP17 ameliorates the impaired NCT in DYT1 MNs. A, Construction and validation of lentiviral vectors expressing GFP-tagged RANBP17 and GFP alone. B, Schematic shows the constructions of lentiviral vectors expressing transport reporters RFP-NLS and RFP-NES and the expected subcellular distribution under normal and impaired NCT conditions. C, Representative confocal images of DYT1 MNs at 21 dpi coexpressed RFP-NLS and GFP or RANBP17-GFP. The nuclei and the soma were highlighted with white and yellow dotted lines, respectively. Scale bar, 50 µm. D, Representative confocal images of DYT1 MNs at 21 dpi coexpressed RFP-NES and GFP or RANBP17-GFP. The nuclei and the soma were highlighted with white and yellow dotted lines, respectively. Scale bar, 50 µm. E, The distribution of RFP-NLS signal under indicated conditions is shown as a ratio of cytoplasmic to nuclear signal. N (neurons) = 120 for health + GFP, 113 for health + RANBP17, 95 for DYT1 + GFP, and 97 for DYT1 + RANBP17 from three independent experiments. ns, no significant difference; **p < 0.01; ***p < 0.001. Student’s t tests. F, The distribution of RFP-NES signal under indicated conditions is shown as a ratio of nuclear to cytoplasmic signal. N (neurons) = 122 for health + GFP, 113 for health + RANBP17, 125 for DYT1 + GFP, and 121 for DYT1 + RANBP17 from three independent experiments. ns, no significant difference; ***p < 0.001, ****p < 0.0001. Student’s t tests. G, The quantification data of FISH assay shown as a ratio of nuclear to cytoplasmic oligo-dT signal. N (neurons) = 104 for health + GFP, 97 for health + RANBP17, 95 for DYT1 + GFP, and 89 for DYT1 + RANBP17 from three independent experiments. ns, no significant difference; **p < 0.01, ****p < 0.0001. Student’s t tests.