Fig. 1.

Potentiated Hsp104 variants rescue toxicity of ALS-linked TDP-43 mutants. (A) Selected strains from B expressing the indicated constructs were induced for 5 h, lysed and then immunoblotted for the specified proteins. A loading control, 3-phosphoglycerate kinase (PGK), was used. (B) Δhsp104 yeast integrated with the indicated pAG303GAL-TDP-43 variant were transformed with the indicated pAG416GAL-Hsp104 variant or vector control. Strains were serially diluted fivefold and spotted on glucose (off) or galactose (on) medium. (C) Fluorescence microscopy of cells co-expressing fluorescently tagged TDP-43 constructs (TDP-43^WT, left; TDP-43^M337V, right) and Hsp104^WT, Hsp104^A503V Hsp104^V426L or vector (416Gal). Cells were stained with DAPI to visualize nuclei (blue). (D) TDP-43 localization was quantified by counting the number of cells containing colocalized nuclear staining. Values represent means±s.e.m. (n=3).