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. 2008 May 19;181(4):655–666. doi: 10.1083/jcb.200712051

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© 2008 Juhász et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 2. — Starvation-induced autophagy in the larval fat body requires Vps34. All images depict live fat body tissue dissected from fed (C) or 4-h starved animals (all other panels). (A) Vps34^Δm22 loss-of-function clones (marked by lack of GFP) fail to accumulate autolysosomes (labeled by punctate Lysotracker Red staining) after starvation. (B) Vps34^KD-expressing cells (GFP positive) fail to accumulate punctate Lysotracker Red staining under starvation conditions. (C and D) Clonal expression of wild-type Vps34 (GFP-positive cells) does not induce autophagy under fed conditions (C) but increases the response to starvation (D). (E) Vps34^Δm22 loss-of-function clones (marked by lack of myrRFP) fail to accumulate autophagosomes (marked by punctate GFP-Atg8) in response to starvation. (F and G) Starvation-induced accumulation of GFP-Atg8a punctae is observed in controls (F) but not in animals expressing Vps34^KD (G). (H–K) TEM images reveal abundant autophagosomes (AP) and autolysosomes (AL) in control animals (H) but not Vps34^Δm22 mutants (I) nor animals expressing Vps34^KD (J). LD, lipid droplet. The graph in K shows autophagosome and autolysosome area ratios calculated from electron micrographs of five animals per genotype. Error bars show SD from the mean. P-values (Mann-Whitney U test): control versus Vps34^Δm22, AP, 1.45e-11 and AL, 3.9e-9; and control versus Vps34^KD, AP, 2.9e-11 and AL, 1.5 e-11. Bars: (A–G) 10 μm; (H–J) 1 μm. Genotypes: (A) hsflp; FRT42D Vps34^Δm22/UAS-2xeGFP FRT42D fb-GAL4, (B) hsflp; Act>CD2>GAL4 UAS-GFP/UAS-Vps34^KD, (C and D) hsflp; Act>CD2>GAL4 UAS-GFP/UAS-Vps34^WT, (E) hsflp; FRT42D Vps34^Δm22/Cg-GAL4 UAS-GFP-Atg8a FRT42D UAS-myrRFP, (F) hsflp; Act>CD2>GAL4 UAS-GFP-Atg8a/+, (G) hsflp; Act>CD2>GAL4 UAS-GFP-Atg8a/UAS-Vps34^KD, (H) Vps34^Δm22/CyO-GFP, (I) Vps34^Δm22/Δm22, (J) Cg-GAL4 UAS-Vps34^KD/Cg-GAL4 UAS-Vps34^KD, and (K) as in H–J.