Figure 5.

Cytoplasmic TDP43-EGFP significantly increases the risk of death in transfected neurons. A–C, Cells displaying purely nuclear (arrow) or both nuclear and cytoplasmic (arrowhead) TDP43-EGFP were identified by fluorescence microscopy. A, EGFP fluorescence. B, mCherry fluorescence. C, Merged image with EGFP fluorescence in green, mCherry fluorescence in red, and overlap in yellow. Scale bar, 10 μm. D, Kaplan-Meier survival analysis of neurons with matched expression levels and either nuclear localization of TDP43-EGFP (nuclear, n = 153) or nuclear and cytoplasmic protein (cytoplasmic, n = 258), demonstrating the toxicity of cytoplasmic TDP43-EGFP. E, The toxic effect of cytoplasmic mislocalization was similar for both WT (n = 170) and mutant TDP43-EGFP (n = 241). *p < 0.0001 (log-rank test). NS, not significant (p > 0.05). Cumulative risk of death curves were constructed with data from three separate experiments.