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. 2022 Feb 11;25(3):103900. doi: 10.1016/j.isci.2022.103900

Figure 4.

Figure 4

MATR3 and MATR3 variants form insoluble species that are subtly resistant to detergent

(A) W303Δhsp104 yeast were transformed with pAG423GAL-MATR3-GFP or the indicated MATR3 mutants. W303WT pAG303GAL-103Q-YFP, W303Δhsp104 pAG303GAL-TDP-43-YFP were used as controls. Strains were induced for 5 h, lysed under native conditions, and analyzed by semi-denaturing detergent agarose gel electrophoresis (SDD-AGE) with the indicated SDS concentrations. MATR3 was detected via immunoblotting against GFP (mouse).

(B) Filter retention assays were performed on the same yeast lysates used in (A) Lysates were applied to cellulose acetate (CA) and nitrocellulose (NC) membranes in parallel and immunoblotted for GFP (rabbit) or PGK.

(C) Quantification of immunoblots from B using 0% SDS samples. The ratio of GFP detection on the CA vs. NC membrane was calculated and normalized to TDP-43-YFP levels. Results were compared to those for the TDP-43-YFP strain using a one-way ANOVA with a Dunnett’s multiple comparisons test (individual data points are shown as dots, bars show means ± SEM, N = 4, ∗p < 0.05).