Figure S1. Trafficking Defects in ESCRT/

(A and B) Coexpression of Vps4DN with P35 in fat body cells results in strong punctuate accumulations of GFP-Atg8a in L2 larvae that are distinct from Hrs-positive endocytic compartments. (C) Hrs-positive endosomes are few and small in fat body cells of a L2 larvae express-ing P35 alone. (D) Eye discs consisting almost entirely of vps25A3 mutant cells accumulate Hrs and Atg8a in distinct punctae. (E) Because there is very little autophagy in wild-type imaginal disc and follicle cells, colocalization between autophagic and en-docytic markers in wild-type cells was in-vestigated in the fat body. As reported in mammalian cells, a subset of GFP-Atg8a structures in mid L3 fat body colocalizes with Hrs (arrows) [S1, S2]. (F and G) GFP-Atg8a structures largely coloc-alize with the late endolysosomal marker HRP-Lamp1 or endocytosed TRD after 20 min pulse and 90 min chase in mid L3 fat body cells. Scale bars represent 10 mm (A), 5 mm (E), and 2 mm (B–D, F, and G). The genotypes are a