Figure 4.
In vivo imaging shows association of lysosomes to trans-Golgi is mostly stable. (A) Setup for in animal imaging of blood cells. An L3 larva is shown immobilized between a glass slide and a coverslip held together by modeling clay. (B) Higher magnification image of the larva in A. (C) Spinning-disc confocal image of live blood cells expressing lysosomal marker Lamp1-GFP (green) and trans-Golgi marker GalT-TagRFP (magenta), both driven by BM-40-SPARC-GAL4. 39 cells were recorded for 10 min each at a speed of 10 images/min. Each image is a maximum intensity projection of a 7–9 section stack (z-distance 1 μm). (D) Still images from a spinning-disc confocal recording of a live blood cell expressing lysosomal marker Lamp1-GFP (green, shown separately on the right) and trans-Golgi marker GalT-TagRFP (magenta). Lysosomes are stably associated with trans-Golgi. Refer to Video 2. (E) Still images from a recording of a blood cell as in D. (F) Still images from a live blood cell as in D. A lysosome (white arrow) is seen detaching from trans-Golgi (magenta arrowhead). Refer to Video 2. (G) Lysosome detaching from trans-Golgi as in F. (H) Still images from a live blood cell as in D. A lysosome (white arrow) is seen attaching to trans-Golgi (magenta arrowhead). Refer to Video 2. (I) Lysosome attaching to trans-Golgi as in H. (J) Still images from a live blood cell as in D. A lysosome (white arrow) is seen detaching from a trans-Golgi element and attaching to a different one (magenta arrowheads). Refer to Video 2. (K) Lysosome detaching from a trans-Golgi element and attaching to a different one as in J. (L) Still images from a live blood cell as in D. A lysosome (white arrow) is seen detaching from a trans-Golgi element (magenta arrowhead) and re-attaching to it. Refer to Video 2.