Nuclear PABPN1 aggregates in the OPMD Drosophila model.

A) Genetic rescue of wing position phenotypes with twin^KG00877 hererozygous mutant. Percentage of wing posture defects in the presence or absence of twin^KG00877/+, scored at day 6. OPMD flies were w¹¹¹⁸; UAS-PABPN1-17ala/+; Mhc-Gal4/+ raised at 18°C. *** p-value <0.0001, using the χ² test. B) Immunostaining of indirect flight muscles from OPMD (w¹¹¹⁸; UAS-PABPN1-17ala/+; Mhc-Gal4/+) and OPMD; twin^KG00877/+ (w¹¹¹⁸; UAS-PABPN1-17ala/+; Mhc-Gal4/twin^KG00877) adult flies at day 6 with anti-PABPN1 (green), showing nuclear aggregates. DNA was visualized with DAPI (blue). Nuclei are outlined with a dotted line. C) Quantification of PABPN1 nuclear aggregates. (Top) Percentages of nuclei with a nuclear PABPN1 aggregate in OPMD and OPMD; twin^KG00877/+ indirect flight muscles (genotypes as in B) *** p-value <0.0001, using the χ² test. (Bottom) Quantification of nuclear aggregate areas. Each nuclear aggregate was delimited in a focal plan and the surface was calculated using ImageJ. Mean values of the areas with standard errors of the mean are indicated in arbitrary units Distribution of cross-sectional areas is shown as box plots (right), the median is indicated as an horizontal line within the box. ns: not significant, using the Student’s t-Test. Thoracic muscles were stained with anti-PABPN1 and DAPI and nuclear aggregates were visualized and scored using both staining. Quantification was from three independent experiments.