Figure 1.
Secretion screening hits Rop, vps20, and VhaPPA1-1 retain cargo inside autophagic vesicles in fat body adipocytes. (A) Schematic drawing of the Drosophila fat body next to a confocal image of L3 adipocytes (feeding stage). In the wild type, Collagen IV is only seen in the basement membrane (BM) that surrounds the tissue and in the equally extracellular CIVICs (Collagen IV intercellular concentrations, circled; Dai et al., 2017). Collagen IV in green (α2 chain Vkg-GFP) and nuclei in blue (DAPI staining). (B) Confocal images of L3 fat body (feeding stage) from wild-type, BM-40-SPARC>Ropi (v19696), >Vps20i (THU5269), and >VhaPPA1-1i (TH04233.N) larvae, showing localization of Collagen IV (α2 chain Vkg-GFP, green). Plasma membrane labeled with BM-40-SPARC-GAL4-driven myr-RFP (magenta). DAPI in blue. Magnified images show accumulation of Collagen IV in large myr-RFP–positive vesicles in mutant conditions. (C) Fat body from wild-type, BM-40-SPARC>Ropi, >Vps20i, and >VhaPPA1-1i L3 larvae showing localization of BM-40-SPARC-GAL4–driven general secretion marker secr-GFP (Wg signal peptide fused to GFP, green). Magnified images show accumulation of secr-GFP in myr-RFP–positive (magenta) vesicles in mutant conditions. (D) Fat body from wild-type, BM-40-SPARC>Ropi, >Vps20i, and >VhaPPA1-1i L3 larvae showing localization of Collagen IV (Vkg-GFP, green) and BM-40-SPARC-GAL4–driven autophagy marker Atg8a-mCherry (magenta). Magnified insets show accumulation of Collagen IV in Atg8a-positive vesicles in mutant conditions. (E) TEM images of autophagic vesicles (defective autolysosomes, inside dashed lines) from L3 fat body (feeding stage) in BM-40-SPARC>Ropi (v19696), >Vps20i (THU5269), and >VhaPPA1-1i (v47188) larvae.

Secretion screening hits Rop, vps20, and VhaPPA1-1 retain cargo inside autophagic vesicles in fat body adipocytes. (A) Schematic drawing of the Drosophila fat body next to a confocal image of L3 adipocytes (feeding stage). In the wild type, Collagen IV is only seen in the basement membrane (BM) that surrounds the tissue and in the equally extracellular CIVICs (Collagen IV intercellular concentrations, circled; Dai et al., 2017). Collagen IV in green (α2 chain Vkg-GFP) and nuclei in blue (DAPI staining). (B) Confocal images of L3 fat body (feeding stage) from wild-type, BM-40-SPARC>Ropi (v19696), >Vps20i (THU5269), and >VhaPPA1-1i (TH04233.N) larvae, showing localization of Collagen IV (α2 chain Vkg-GFP, green). Plasma membrane labeled with BM-40-SPARC-GAL4-driven myr-RFP (magenta). DAPI in blue. Magnified images show accumulation of Collagen IV in large myr-RFP–positive vesicles in mutant conditions. (C) Fat body from wild-type, BM-40-SPARC>Ropi, >Vps20i, and >VhaPPA1-1i L3 larvae showing localization of BM-40-SPARC-GAL4–driven general secretion marker secr-GFP (Wg signal peptide fused to GFP, green). Magnified images show accumulation of secr-GFP in myr-RFP–positive (magenta) vesicles in mutant conditions. (D) Fat body from wild-type, BM-40-SPARC>Ropi, >Vps20i, and >VhaPPA1-1i L3 larvae showing localization of Collagen IV (Vkg-GFP, green) and BM-40-SPARC-GAL4–driven autophagy marker Atg8a-mCherry (magenta). Magnified insets show accumulation of Collagen IV in Atg8a-positive vesicles in mutant conditions. (E) TEM images of autophagic vesicles (defective autolysosomes, inside dashed lines) from L3 fat body (feeding stage) in BM-40-SPARC>Ropi (v19696), >Vps20i (THU5269), and >VhaPPA1-1i (v47188) larvae.

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