(A) NBD1 variants suppress TDP-43 toxicity in yeast. W303aΔhsp104-pAG-303GAL-TDP-43 yeast were transformed with Hsp104 variants or vector. Strains were serially diluted 5-fold and spotted in duplicate onto glucose (non-inducing) and galactose (inducing) media.
(B) NBD1 variants do not reduce TDP-43 expression in yeast. Strains in (A) were induced for 5 h, lysed, and immunoblotted for Hsp104, TDP-43, and PGK (loading control).
(C) NBD1 variants suppress TDP-43 aggregation and mislocalization in yeast. Fluorescence microscopy of cells coexpressing fluorescently tagged TDP-43 and Hsp104 variants. Strains were induced for 5 h in galactose, fixed, and stained with DAPI (blue) to visualize nuclei. Scale bar, 2.5 μm.
(D) TDP-43 localization was quantified by calculating the proportion of cells containing colocalized nuclear staining. Values represent means ± SEM (n = 3).
See also Figures S1 and S2.