High-resolution analysis of tGA-lysosomes. (A) Superresolution (SIM) images of fat body ERES-Golgi units showing trans-Golgi localization of lysosomal marker Lamp1-GFP (green). GalT-TagRFP (trans-Golgi, magenta) and ManII-BFP (mid-Golgi, blue) are shown as well, all driven by BM-40-SPARC-GAL4. (B) Superresolution (SIM) images of fat body ERES-Golgi units showing trans-Golgi localization of recycling endosome marker Rab11-GFP (green). GalT-TagRFP (trans-Golgi, magenta) and ManII-BFP (mid-Golgi, blue) are shown as well, BM-40-SPARC-GAL4. (C) FIB-SEM image of a fat body (FB) ERES-Golgi unit displaying a tGA-lysosome. The same image is pseudocolored on the right side in blue (ERES), purple (Golgi), and yellow (tGA-lysosome). The black staining marks the localization between ERES and Golgi of γCOP (Cg-GAL4-driven γCOP-APEX). (D) FIB-SEM images featuring examples of tGA-lysosomes in fat body. Superimposed colors in lower images indicate Golgi (purple) and tGA-lysosome (yellow). (E) FIB-SEM images featuring examples of tGA-lysosomes in imaginal wing disc (WD). Superimposed colors in lower images as in B. (F) FIB-SEM image (left) and reconstructed 3D model (right) of a tGA-lysosome (yellow) sitting on Golgi (purple). Refer to Video 1. (G) FIB-SEM image and reconstructed 3D model of a tGA-lysosome (yellow) standing a short distance from Golgi (purple). Refer to Video 1. (H) FIB-SEM serial images (left, 20 nm Z-distance) and reconstructed 3D model (right) of a tGA-lysosome connected (arrow) to Golgi. Refer to Video 1. (I) FIB-SEM serial images (20 nm Z-distance) and reconstructed 3D model of a tGA-Golgi lysosome connected (arrows) to Golgi. Refer to Video 1.