Figure 3.

Stabilized Acn^D527A enhances basal autophagy. (A and B) Micrographs of fed Acn^wt or Acn^D527A larval eye discs stained for Atg8a. (C) Quantification of Atg8a punctae in eye discs from five larvae. (D) Western blot of fat body lysates from 96-h fed larvae probed for Acn and Actin. (E) Quantification of blots for Acn levels relative to Actin (n = 3). (F–G′ and I–J′) Micrographs of Acn^wt or Acn^D527A fat bodies encompassing six to eight cells from 96-h fed or starved size-matched larvae. (F–G′) Fat bodies stained with antibodies against Atg8a. (H) Quantification of Atg8a punctae averaged from five larvae from one representative experiment out of three repeats. (I–J′) GFP-Atg8a fluorescence detected in live fat bodies. (K) Quantification of GFP-Atg8a punctae averaged from five larvae from one representative experiment out of three repeats. (L–M″) TEMs of fed Acn^wt or Acn^D527A fat bodies. Examples of dense lysosomes (arrowheads) and membrane-enriched autolysosomes (arrows) are shown at higher magnification. (N) Quantification of percentage of autolysosomal area averaged from 25 images per genotype. (O) Quantification of diameters of ≥100 lysosomes and autolysosomes per genotype. Detailed genotypes are given in Table S2. wt, wild type. Error bars show means ± SD. **, P < 0.01; ***, P < 0.001.