Figure 2.

MATR3 forms round nuclear puncta in yeast, while some MATR3 mutants form misshapen puncta

(A) W303Δhsp104 yeast were transformed with the indicated pAG423GAL-MATR3-GFP variant or vector control. Strains were serially diluted 5-fold and spotted in duplicate on glucose (non-inducing) and galactose (inducing) media. Scores are based on number of spots from an average of at least three biological replicates.

(B) Strains from A were induced for 5 h, lysed, and immunoblotted for GFP (mouse) and PGK (loading control).

(C) Strains from A were induced for 5 h, stained with Hoechst dye, and imaged. Representative images are shown. Merge channel includes GFP, Hoechst, and bright field images. Scale bar, 5μm.

(D) Strains from A were imaged at higher resolution using a confocal microscope with Airyscan deconvolution (Airyscan parameter 2.7). Representative images are shown. Scale bar, 1μm.

(E) Circularity ratio of foci of MATR3-GFP (N = 233 cells), MATR3:S85C-GFP (N = 241 cells), MATR3:F115C-GFP (N = 252 cells), MATR3:P154S-GFP (N = 167 cells), MATR3:T622A-GFP (N = 245 cells) from experiments in D. Results were compared to those for the MATR3-GFP strain using a one-way ANOVA with a Dunnett’s multiple comparisons test (individual data points are shown as dots, bars show means, ∗∗p < 0.005, ∗∗∗∗p < 0.0001).