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. 2018 Jan 2;217(1):361–374. doi: 10.1083/jcb.201702145

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© 2018 Csizmadia et al.

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Figure 1. — Time course of developmentally programmed crinophagy in Drosophila salivary gland cells. (A–D) Glue granule degradation in the salivary gland of animals coexpressing Glue-GFP and Glue-Red reporters (GlueFlux). (A) Wandering L3 stage (−6 h RPF) larval cells contain mostly intact (GFP and DsRed double-positive) secretory granules, and very few degrading glue granules (positive for DsRed only) are seen. (B–D) The number of intact (double positive) glue granules gradually decreases in −2-h (B), 0-h (C), and 4-h (D) old animals, in parallel with the increasing number of DsRed-only crinosomes. (E–H) Glue granules acquire lysosomal Cathepsin B (CathB) 3xmCherry. (E) No colocalization is observed between Glue-GFP granules and CathB at −6 h. (F) CathB structures greatly increase in size and number, and many overlap with Glue-GFP granules (arrowheads) at −2 h. Note that GFP fluorescence often decreases in overlapping structures. (G) The majority of Glue-GFP granules are positive for CathB at 0 h. (H) GFP fluorescence is lost in most granules at 4 h, and the size and number of CathB structures resembles that of Glue-Red in D. (I and J) Glue granule fusion with late endosomes and lysosomes. (I) GFP-Lamp1 localizes to small punctate structures in wandering-stage larvae and very rarely forms rings around Glue-Red granules. (J) GFP-Lamp1 forms prominent rings around Glue-Red granules at −2 h, likely as a result of secretory granule fusion with late endosomes and lysosomes. The boxed regions in I and J are shown enlarged in I′ and J′, with arrowheads pointing to GFP-Lamp1 rings. Green and/or magenta channels of merged images are also shown separately as indicated. Bars: (A–J) 20 µm; (I′ and J′) 3 µm. Please see Fig. S1 (A, F, and G) for quantification of data in A–D, E–H, I, and J, respectively.