Basal autophagy takes place in the trans-Golgi–associated degradative compartment under homeostatic conditions. (A) Confocal images of L3 fat body (feeding stage) from wild-type (left) and BM-40-SPARC>Atg8aⁱ (right) larvae showing localization of autophagic cargo Ref(2)P (antibody staining, green) and trans-Golgi marker GalT-TagRFP (magenta, driven by BM-40-SPARC-GAL4). Examples of Ref(2)P puncta proximal to trans-Golgi are shown at higher magnification. Nuclei in blue (DAPI). (B) Proximity between Ref(2)P and GalT-TagRFP in wild-type fat body. The upper graph represents proximity measured in 10 cells. The lower graph represents average proximity values (n = 10, error bars indicate SD). (C) Localization of Collagen IV (α2 chain Vkg-GFP) in L3 fat body (feeding stage) from wild type (left), BM-40-SPARC>Atg8aⁱ (center left), and wild type treated for 12 h with autophagy inhibitors Bafilomycin A (2 μM, center right) and chloroquine (15 mg/ml, right). Collagen IV puncta in the wild type correspond not to intracellular accumulation, but to CIVICs (see Fig. 1 A). BM indicates basement membrane.