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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 1. — Reagents to disrupt and monitor Vps34 function in D. melanogaster. (A) Map of the Vps34 locus, showing the location and extent of the Δm22 deletion. (B) RT-PCR analysis of Vps34 mRNA expression in control and mutant third instar larvae. Vps34 transcript levels are strongly reduced in ex32 animals and undetectable in the Δm22 mutant. (C–E) Disruption of Vps34 function causes mislocalization of the PI(3)P reporter GFP-2xFYVE. In wild-type fat body cells (C and D), GFP-2xFYVE accumulates at the perinuclear endosomal compartment. This pattern of localization is lost in Vps34^Δm22 clones (C; mutant cell marked by lack of dsRed) or in response to expression of Vps34^KD (E). Bar, 10 μm. Genotypes: (C) hsflp; FRT42D UAS-GFP-2xFYVE Vps34^Δm22/Cg-GAL4 FRT42D UAS-GFP-2xFYVE UAS-dsRed, (D) hsflp; UAS-GFP-2xFYVE/+; Act>CD2>GAL4/+, and (E) hsflp; UAS-GFP-2xFYVE/+ ; Act>CD2>GAL4 / UAS-Vps34^KD.