Figure 4.

Rapamycin Increases Autophagy and Decreases Protein Translation

(A) Levels of ³⁵S-methionine incorporation in 7-day-old flies normalized to total protein content (±SEM). Rapamycin significantly reduces ³⁵S-methionine incorporation (^∗∗p = 0.0033, Student's t test).

(B) Significantly more autolysosomes and lysosomes in rapamycin-treated flies (^∗∗∗p < 0.0001, Student's t test). Average number of LysoTracker-stained puncta in fly midguts isolated from rapamycin pretreated or control flies is presented (±SEM).

(C) Representative confocal fluorescence images of fly midgut stained with LysoTracker Red. Midgut of control flies (upper image) and rapamycin pretreated flies (lower image) are shown. L indicates lumen of the gut; scale bar is 10 μM.