Fig. 5.
Partial suppression of locomotor deficits, muscle degeneration and polyubiquitin aggregate accumulation by decreasing Tor activity. All animals analyzed were males that had been reared as mating pairs. (A) elav>dcr2,atlRNAi flies (solid circles) or Tork17004/+; elav>dcr2,atlRNAi flies (open circles) that had been maintained at 25°C as single mating pairs were analyzed for viability (blue and red circles) and paralysis (green circles). Mean±s.e.m. of the percentage of flies living, paralyzed or dead from seven replicates with 70 flies for Tork17004/+; elav>dcr2,atlRNAi and 73 flies for elav>dcr2,atlRNAi is shown. (B–I) Dissected thoracic tissue from 1-day-old (B–E) or 7-day-old (F–I) elav>dcr2,atlRNAi (B,D,F and H) or Tork17004/+;elav>dcr2,atlRNAi (C,E,G and I) adults was fixed and stained with Alexa-Fluor-647–phalloidin (B,C,F and G) or an anti-ubiquitin antibody (D,E,H and I). DFM49 was visualized. Scale bars: 25 µm. Each image is a single slice from a confocal z-stack. (J–K) 0-day-old (J) or 7-day-old (K) Tork17004/+;elav>dcr2,atlRNAi males (blue circles represent each individual animal DFM49) or elav>dcr2,atlRNAi males (green circles represent each individual animal DFM49) were dissected and analyzed. Alexa-Fluor-647–phalloidin staining was used to outline DFM49. (J) Quantification of polyubiquitin pixel intensities per µm3 in each muscle following staining with an anti-ubiquitin antibody. Means±s.e.m. are indicated. P=0.024 by Student's t-test for elav>dcr2,atlRNAi flies versus Tork17004/+;elav>dcr2,atlRNAi. (K) Quantification of actin levels by measuring Alexa-Fluor-647 pixel intensities per µm3 following staining with Alexa-Fluor-647–phalloidin. Means±s.e.m. are indicated. P=0.003 by Student's t-test for elav>dcr2,atlRNAi flies versus Tork17004/+;elav>dcr2,atlRNAi.