Bäumers, M., Klose, S., Brüser, C., Haag, C., Hänsch, S., Pannen, H., Weidtkamp-Peters, S., Feldbrügge, M., & Klein, T. (2019). The auxiliary ESCRT complexes provide robustness to cold in poikilothermic organisms. *Biology Open*, 8, bio043422. https://doi.org/10.1242/bio.043422

Becker, A., Thakur, B. K., Weiss, J. M., Kim, H. S., Peinado, H., & Lyden, D. (2016). Extracellular vesicles in cancer: Cell-to-cell mediators of metastasis. *Cancer Cell*, 30, 836–848. https://doi.org/10.1016/j.ccell.2016.10.009

Bras, S. L., Rondanino, C., Kriegel-Taki, G., Dussert, A., & Borgne, R. L. (2012). Genetic identification of intracellular trafficking regulators involved in Notch-dependent binary cell fate acquisition following asymmetric cell division. *Journal of Cell Science*, 125, 4886–4901. https://doi.org/10.1242/jcs.110171

Carlton, J. G., Agromayor, M., & Martin-Serrano, J. (2008). Differential requirements for Alix and ESCRT-III in cytokinesis and HIV-1 release. *Proceedings of the National Academy of Sciences*, 105, 10541–10546. https://doi.org/10.1073/pnas.0802008105

Colombo, M., Raposo, G., & Théry, C. (2014). Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. *Annual Review of Cell and Developmental Biology*, 30, 255–289. https://doi.org/10.1146/annurev-cellbio-101512-122326

Comito, G., Ippolito, L., Chiarugi, P., & Cirri, P. (2020). Nutritional exchanges within tumor microenvironment: Impact for cancer aggressiveness. *Frontiers in Oncology*, 10, 396. https://doi.org/10.3389/fonc.2020.00396

Corrigan, L., Redhai, S., Leiblich, A., Fan, S.-J., Perera, S. M. W., Patel, R., Gandy, C., Wainwright, S. M., Morris, J. F., Hamdy, F., Goberdhan, D. C. I., & Wilson, C. (2014). BMP-regulated exosomes from Drosophila male reproductive glands reprogram female behavior. *Journal of Cell Biology*, 206, 671–688. https://doi.org/10.1083/jcb.201401072

Dar, G. H., Mendes, C. C., Kuan, W.-L., Speciale, A. A., Conceição, M., Görgens, A., Uliyakina, I., Lobo, M. J., Lim, W. F., El-Andaloussi, S., Mager, I., Roberts, T. C., Barker, R. A., Goberdhan, D. C. I., Wilson, C., & Wood, M. J. A. (2021). GAPDH controls extracellular vesicle biogenesis and enhances the therapeutic potential of EV mediated siRNA delivery to the brain. *Nature Communications*, 12, 6666. https://doi.org/10.1038/s41467-021-27056-3

Dietzl, G., Chen, D., Schnorrer, F., Su, K.-C., Barinova, Y., Fellner, M., Gasser, B., Kinsey, K., Oppel, S., Scheiblauer, S., Couto, A., Marra, V., Keleman, K., & Dickson, B. J. (2007). A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. *Nature*, 448, 151–156. https://doi.org/10.1038/nature05954

Dimaano, C., Jones, C. B., Hanono, A., Curtiss, M., & Babst, M. (2008). Ist1 regulates Vps4 localization and assembly. *Molecular Biology of the Cell*, 19, 465–474. https://doi.org/10.1091/mbc.E07-08-0747

Dunst, S., Kazimiers, T., von Zadow, F., Jambor, H., Sagner, A., Brankatschk, B., Mahmoud, A., Spannl, S., Tomancak, P., Eaton, S., & Brankatschk, M. (2015). Endogenously tagged Rab proteins: A resource to study membrane trafficking in Drosophila. *Developmental Cell*, 33, 351–365. https://doi.org/10.1016/j.devcel.2015.03.022

Fan, S.-J., Kroeger, B., Marie, P. P., Bridges, E. M., Mason, J. D., McCormick, K., Zois, C. E., Sheldon, H., Khalid Alham, N., Johnson, E., Ellis, M., Stefana, M. I., Mendes, C. C., Wainwright, S. M., Cunningham, C., Hamdy, F. C., Morris, J. F., Harris, A. L., Wilson, C., ... Goberdhan, D. C. (2020). Glutamine deprivation alters the origin and function of cancer cell exosomes. *Embo Journal*, 39, e103009. https://doi.org/10.15252/embj.2019103009

Fan, S. J., Snell, C., Turley, H., Li, J. L., McCormick, R., Perera, S. M., Heublein, S., Kazi, S., Azad, A., Wilson, C., Harris, A. L., & Goberdhan, D. C. (2016). PAT4 levels control amino-acid sensitivity of rapamycin-resistant mTORC1 from the Golgi and affect clinical outcome in colorectal cancer. *Oncogene*, 35, 3004–3015. https://doi.org/10.1038/onc.2015.363

Frankel, E. B., Shankar, R., Moresco, J. J., Yates, J. R., Volkmann, N., & Audhya, A. (2017). Ist1 regulates ESCRT-III assembly and function during multivesicular endosome biogenesis in *Caenorhabditis elegans* embryos. *Nature Communications*, 8, 14390. https://doi.org/10.1038/s41467-017-01636-8

Gomez-Lamarca, M. J., Snowdon, L. A., Seib, E., Klein, T., & Bray, S. J. (2015). Rme-8 depletion perturbs Notch recycling and predisposes to pathogenic signaling. *Journal of Cell Biology*, 210, 303–318. https://doi.org/10.1083/jcb.201411001

Gondré-Lewis, M. C., Park, J. J., & Loh, Y. P. (2012). Cellular mechanisms for the biogenesis and transport of synaptic and dense-core vesicles. *International Review of Cell and Molecular Biology*, 299, 27–115. https://doi.org/10.1016/B978-0-12-394310-1.00002-3

Henne, W. M., Buchkovich, N. J., & Emr, S. D. (2011). The ESCRT pathway. *Developmental Cell*, 21, 77–91. https://doi.org/10.1016/j.devcel.2011.05.015

Henne, W. M., Buchkovich, N. J., Zhao, Y., & Emr, S. D. (2012). The endosomal sorting complex ESCRT-II mediates the assembly and architecture of ESCRT-III helices. *Cell*, 151, 356–371. https://doi.org/10.1016/j.cell.2012.08.039

Hochstrasser, M. (2009). Origin and function of ubiquitin-like proteins. *Nature*, 458, 422–429. https://doi.org/10.1038/nature07958

Irion, U., & St Johnston, D. (2007). bicoid RNA localization requires specific binding of an endosomal sorting complex. *Nature*, 445, 554–558. https://doi.org/10.1038/nature05503

Jiang, H., Patel, P. H., Kohlmaier, A., Grenley, M. O., McEwen, D. G., & Edgar, B. A. (2009). Cytokine/Jak/Stat signaling mediates regeneration and homeostasis in the *Drosophila* midgut. *Cell*, 137, 1343–1355. https://doi.org/10.1016/j.cell.2009.05.014

König, J., Frankel, E. B., Audhya, A., & Müller-Reichert, T. (2017). Membrane remodeling during embryonic abscission in *Caenorhabditis elegans*. *Journal of Cell Biology*, 216, 1277–1286. https://doi.org/10.1083/jcb.201607030

Kubli, E. (2003). Sex-peptides: Seminal peptides of the *Drosophila* male. *Cellular and Molecular Life Sciences*, 60, 1689–1704. https://doi.org/10.1007/s00018-003-3052

Lee, J. -A., Beigneux, A., Ahmad, S. T., Young, S. G., & Gao, F.-B. (2007). ESCRT-III dysfunction causes autophagosome accumulation and neurodegeneration. *Current Biology*, 17, 1561–1567. https://doi.org/10.1016/j.cub.2007.07.029

Leiblich, A., Marsden, L., Gandy, C., Corrigan, L., Jenkins, R., Hamdy, F., & Wilson, C. (2012). Bone morphogenetic protein- and mating-dependent secretory cell growth and migration in the Drosophila accessory gland. *Proc Natl Acad Sci U S A.*, 109, 19292–19297. https://doi.org/10.1073/pnas.1214517109

Li, J., Belogortseva, N., Porter, D., & Park, M. (2008). Chmp1A functions as a novel tumor suppressor gene in human embryonic kidney and ductal pancreatic tumor cells. *Cell Cycle Georget. Tex*, 7, 2886–2893. https://doi.org/10.4161/cc.7.18.6677

Li, X., Suo, J., Shao, S., Xue, L., Chen, W., Dong, L., Shi, J., Fu, M., Lu, N., Zhan, Q., & Tong, T. (2014). Overexpression of OLC1 promotes tumorigenesis of human esophageal squamous cell carcinoma. *PLoS ONE*, 9, e90958. https://doi.org/10.1371/journal.pone.0090958

Liu, J., Song, H., Yao, L., Liu, Y., Zhang, Y., Zhao, H., Ji, H., & Wang, Y. (2014). Over-expression of the overexpressed in lung cancer-1 is associated with poor prognosis in colorectal cancer. *Anticancer Research*, 34, 367–372.

Loncle, N., Agromayor, M., Martin-Serrano, J., & Williams, D. W. (2015). An ESCRT module is required for neuron pruning. *Scientific Reports*, 5, 8461. https://doi.org/10.1038/srep08461

Mamińska, A., Bartosik, A., Banach-Orłowska, M., Pilecka, I., Jastrzębski, K., Zdżalik-Bielecka, D., Castanon, I., Poulain, M., Neyen, C., Wolińska-Nizioł, L., Toruń, A., Szymańska, E., Kowalczyk, A., Piwocka, K., Simonsen, A., Stenmark, H., Fürthauer, M., González-Gaitán, M., & Miaczynska, M. (2016). ESCRT proteins restrict constitutive NF-κB signaling by trafficking cytokine receptors. *Science Signaling*, 9, ra8–ra8. https://doi.org/10.1126/scisignal.aad0848

Matias, N. R., Mathieu, J., & Huynh, J. - R. (2015). Abscission is regulated by the ESCRT-III protein shrub in drosophila germline stem cells. *Plos Genetics*, 11, e1004653. https://doi.org/10.1371/journal.pgen.1004653
Matusek, T., Théron, P., & Fürthauer, M. (2019). Functional analysis of ESCRT-positive extracellular vesicles in the *Drosophila* wing imaginal disc. *Methods Molecular Biology*, **1998**, *31*–47. https://doi.org/10.1007/978-1-4939-9492-2_3

Matusek, T., Wendler, F., Polès, S., Pizette, S., D'Angelo, G., Fürthauer, M., & Théron, P. P. (2014). The ESCRT machinery regulates the secretion and long-range activity of Hedgehog. *Nature*, **516**, 99–103. https://doi.org/10.1038/nature13847

McAlister, G. C., Nusinow, D. P., Jedrychowski, M. P., Wühr, M., Huttlin, E. L., Erickson, B. K., Rad, R., Haas, W., & Gygi, S. P. (2014). MultiNotch MS3 enables accurate, sensitive, and multiplexed detection of differential expression across cancer cell line proteomes. *Analytical Chemistry*, **86**, 7150–7158. https://doi.org/10.1021/ac502040v

McCullough, J., Clippinger, A. K., Talledge, N., Skowyra, M. L., Saunders, M. G., Naismith, T. V., Colf, L. A., Afonine, P., Arthur, C., Sundquist, W. I., Hanson, P. I., & Frost, A. (2015). Structure and membrane remodeling activity of ESCRT-III helical polymers. *Science*, **350**, 1548–1551. https://doi.org/10.1126/science.aad8305

Mizuno, E., Kobayashi, K., Yamamoto, A., Kitamura, N., & Komada, M. (2006). A deubiquitinating enzyme UBPY regulates the level of protein ubiquitination on endosomes. *Traffic*, **7**, 1017–1031. https://doi.org/10.1111/j.1600-0854.2006.00452.x

Möbius, W., Ohno-Iwashita, Y., Donselaar, E. G., Oorschot, V. M. J., Shimada, Y., Fujimoto, T., Heijnen, H. F. G., Geuze, H. J., & Slot, J. W. (2002). Immunoelectron microscopic localization of cholesterol using biotinylated and non-cytolytic Perfringolysin O. *Journal of Histochemistry and Cytochemistry*, **50**, 43–55. https://doi.org/10.1177/002215540205000105

Mochida, G. H., Ganesh, V. S., de Michelena, M. I., Dias, H., Atabay, K. D., Kathrein, K. L., Huang, H.-T., Hill, R. S., Felie, J. M., Rakiec, D., Gleason, D., Hill, A. D., Malik, A. N., Barry, B. J., Partlow, J. N., Tan, W.-H., Glader, L. J., Barkovich, A. J., Dobyns, W. B., ... Walsh, C. A. (2012). CHMPIA encodes an essential regulator of BMI1-INK4A in cerebellar development. *Nature Genetics*, **44**, 1260–1264. https://doi.org/10.1038/ng.2425

Neyen, C., Runchel, C., Schüpfer, F., Meier, P., & Lemaitre, B. (2016). The regulatory isoform rPGRP-LC induces immune resolution via endosomal degradation of receptors. *Nature Immunology*, **17**, 1150–1158. https://doi.org/10.1038/ni.3536

Ni, J.-Q., Liu, L.-P., Binari, R., Hardy, R., Shim, H.-S., Cavallaro, A., Booker, M., Pfeiffer, B. D., Markstein, M., Wang, H., Villalta, C., Laverty, T. R., Perkins, L. A., & Perrimon, N. (2009). A Drosophila resource of transgenic RNAi lines for neurogenetics. *Genetics*, **182**, 1089–1100. https://doi.org/10.1534/genetics.109.10360

Nickerson, D. P., West, M., Henry, R., & Odorizzi, G. (2010). Regulators of Vps4 ATPase activity at endosomes differentially influence the size and rate of formation of intraluminal vesicles. *Molecular Biology of the Cell*, **21**, 1023–1032. https://doi.org/10.1091/mbc.E09-09-0776

Ou-Yang, Q. - H., Duan, Z. - X., Jin, Z., & Lei, J. - X. (2014). OLC1 is overexpressed in breast cancer and its expression correlates with poor patient survival. *Tumour Tumour Biology*, **35**, 8823–8827. https://doi.org/10.1007/s13277-014-2130-7

Prag, G., Watson, H., Kim, Y. C., Beach, B. M., Ghirlando, R., Hummer, G., Bonifacino, J. S., & Hurley, J. H. (2007). The Vps27/Hsel complex is a GAT domain-based scaffold for ubiquitin-dependent sorting. *Developmental Cell*, **12**, 973–986. https://doi.org/10.1016/j.devcel.2007.04.013

Prince, E., Kroeger, B., Gligorov, D., Wilson, C., Eaton, S., Karch, F., Brankatschk, M., & Maeda, R. K. (2019). Rab-mediated trafficking in the secondary cells of Drosophila male accessory glands and its role in fecundity. *Traffic*, **20**, 137–151. https://doi.org/10.1111/tra.12622

Raiborg, C., Bache, K. G., Mehlum, A., Stang, E., & Stenmark, H. (2001). Hrs recruits clathrin to early endosomes. *Embo Journal*, **20**, 5008–5021. https://doi.org/10.1093/emboj/20.17.5008

Raymond, C. K., Howald-Stevenson, I., Vater, C. A., & Stevens, T. H. (1992). Morphological classification of the yeast vacuolar protein sorting mutants: Evidence for a prevacuolar compartment in class E vps mutants. *Molecular Biology of the Cell*, **3**, 1389–1402.

Razi, M., & Futter, C. E. (2006). Distinct roles for Tsg101 and Hrs in multivesicular body formation and inward vesiculation. *Molecular Biology of the Cell*, **17**, 3469–3483. https://doi.org/10.1091/mbc.E05-11-1054

Redhai, S., Hellberg, J. E., Wainwright, M., Perera, S. W., Castellanos, F., Kroeger, B., Gandy, C., Leiblich, A., Corrigan, L., Hilton, T., Patel, B., Fan, S. J., Hamdy, F., Goberdhan, D. C., & Wilson, C. (2016). Regulation of dense-core granule replenishment by autocrine BMP signalling in *Drosophila* secondary cells. *Plos Genetics*, **12**, e1006366. https://doi.org/10.1371/journal.pgen.1006366

Rideout, E. J., Dornan, A. J., Neville, M. C., Eadie, S., & Goodwin, S. F. (2010). Control of sexual differentiation and behavior by the doublesex gene in Drosophila melanogaster. *Nature Neuroscience*, **13**, 458–466. https://doi.org/10.1038/nn.2515

Rieder, S. E., Banta, L. M., Köhrer, K., McCaffery, J. M., & Emr, S. D. (1996). Multilamellar endosome-like compartment accumulates in the yeast vps28 vacuolar protein sorting mutant. *Molecular Biology of the Cell*, **7**, 985–999. https://doi.org/10.1091/mbc.7.6.985

Rue, S. M., Mattei, S., Saksena, S., & Emr, S. D. (2007). Novel Ist1-Did2 complex functions at a late step in multivesicular body sorting. *Molecular Biology of the Cell*, **19**, 475–484. https://doi.org/10.1091/mbc.e07-07-0694

Rusten, T. E., Vaccari, T., Lindmo, K., Rodahl, L. M. W., Nezis, I. P., Sem-Jacobsen, C., Wendler, F., Vincent, J.-P., Brech, A., Bilder, D., & Stenmark, H. (2007). ESCRTs and Fab1 regulate distinct steps of autophagy. *Current Biology*, **17**, 1817–1825. https://doi.org/10.1016/j.cub.2007.09.032

Saber, S. H., Ali, H. E. A., Gaballa, R., Gaballah, M., Ali, H. I., Zerfaoui, M., & Abd Elmageed, Z. Y. (2020). Exosomes are the driving force in preparing the soil for the metastatic seeds: Lessons from the prostate cancer. *Cells*, **9**, 564. https://doi.org/10.3390/cells9030564

Saksena, S., & Emr, S. D. (2009). ESCRTs and human disease. *Biochemical Society Transactions*, **37**, 167–172. https://doi.org/10.1042/BST0370167

Schindelin, J., Rueden, C. T., Hiner, M. C., & Eliceiri, K. W. (2015). The ImageJ ecosystem: An open platform for biomedical image analysis: T HE I MAGEJ E COSYSTEM. *Molecular Reproduction and Development*, **82**, 518–529. https://doi.org/10.1002/mrd.22489

Schuh, A. L., & Audhya, A. (2014). The ESCRT machinery: From the plasma membrane to endosomes and back again. *Critical Reviews in Biochemistry and Molecular Biology*, **49**, 242–261. https://doi.org/10.3109/10409238.2014.881777

Sheng, Z., Yu, L., Zhang, T., Pei, X., Li, X., Zhang, Z., & Du, W. (2016). ESCRT-0 complex modulates Rbf-mutant cell survival by regulating Rhomboid endosomal trafficking and EGFR signaling. *Journal of Cell Science*, **129**, 2075–2084. https://doi.org/10.1242/jcs.182261

Shiflett, S. L., Ward, D. M., Huynh, D., Vaughn, M. B., Simmons, J. C., & Kaplan, J. (2004). Characterization of Vtalp, a Class E Vps protein in *Saccharomyces cerevisiae*. *Journal of Biological Chemistry*, **279**, 10982–10990. https://doi.org/10.1074/jbc.M312669200

Shim, J. - H., Xiao, C., Hayden, M. S., Lee, K. - Y., Trombetta, E. S., Pypaert, M., Nara, A., Yoshimori, T., Wilm, B., Erdjument-Bromage, H., Tempst, P., Hogan, B. L. M., Mellman, I., & Ghosh, S. (2006). CHMP5 is essential for late endosome function and down-regulation of receptor signaling during mouse embryogenesis. *Journal of Cell Biology*, **172**, 1045–1056. https://doi.org/10.1083/jcb.200509041

Stuffers, S., Wegner, C. S., Stenmark, H., & Brech, A. (2009). Multivesicular endosome biogenesis in the absence of ESCRTs. *Traffic*, **10**, 925–937. https://doi.org/10.1111/j.1600-0854.2009.00920.x

Teis, D., Saksena, S., Judson, B. L., & Emr, S. D. (2010). ESCRT-II coordinates the assembly of ESCRT-III filaments for cargo sorting and multivesicular body vesicle formation. *Embo Journal*, **29**, 871–883. https://doi.org/10.1038/emboj.2009.408

Théry, C., Witwer, K. W., Aikawa, E., Alcaraz, M. J., Anderson, J. D., Andriantsitohaina, R., Antoniou, A., Arab, T., Archer, F., Atkin-Smith, G. K., Ayre, D. C., Bach, J.-M., Bachurski, D., Baharvand, H., Balaj, L., Baldacchino, S., Bauer, N. N., Baxter, A. A., Bebawy, M., ... Zuba-Surma, E. K. (2018). Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. *Journal of Extracellular Vesicles*, **7**, 1535750. https://doi.org/10.1080/20013078.2018.1535750
Matusek, T., Théron, P., & Fürthauer, M. (2019). Functional analysis of ESCRT-positive extracellular vesicles in the *Drosophila* wing imaginal disc. *Methods Molecular Biology*, **1998**, *31*–47. https://doi.org/10.1007/978-1-4939-9492-2_3

Matusek, T., Wendler, F., Polès, S., Pizette, S., D'Angelo, G., Fürthauer, M., & Théron, P. P. (2014). The ESCRT machinery regulates the secretion and long-range activity of Hedgehog. *Nature*, **516**, 99–103. https://doi.org/10.1038/nature13847

McAlister, G. C., Nusinow, D. P., Jedrychowski, M. P., Wühr, M., Huttlin, E. L., Erickson, B. K., Rad, R., Haas, W., & Gygi, S. P. (2014). MultiNotch MS3 enables accurate, sensitive, and multiplexed detection of differential expression across cancer cell line proteomes. *Analytical Chemistry*, **86**, 7150–7158. https://doi.org/10.1021/ac502040v

McCullough, J., Clippinger, A. K., Talledge, N., Skowyra, M. L., Saunders, M. G., Naismith, T. V., Colf, L. A., Afonine, P., Arthur, C., Sundquist, W. I., Hanson, P. I., & Frost, A. (2015). Structure and membrane remodeling activity of ESCRT-III helical polymers. *Science*, **350**, 1548–1551. https://doi.org/10.1126/science.aad8305

Mizuno, E., Kobayashi, K., Yamamoto, A., Kitamura, N., & Komada, M. (2006). A deubiquitinating enzyme UBPY regulates the level of protein ubiquitination on endosomes. *Traffic*, **7**, 1017–1031. https://doi.org/10.1111/j.1600-0854.2006.00452.x

Möbius, W., Ohno-Iwashita, Y., Donselaar, E. G., Oorschot, V. M. J., Shimada, Y., Fujimoto, T., Heijnen, H. F. G., Geuze, H. J., & Slot, J. W. (2002). Immunoelectron microscopic localization of cholesterol using biotinylated and non-cytolytic Perfringolysin O. *Journal of Histochemistry and Cytochemistry*, **50**, 43–55. https://doi.org/10.1177/002215540205000105

Mochida, G. H., Ganesh, V. S., de Michelena, M. I., Dias, H., Atabay, K. D., Kathrein, K. L., Huang, H.-T., Hill, R. S., Felie, J. M., Rakiec, D., Gleason, D., Hill, A. D., Malik, A. N., Barry, B. J., Partlow, J. N., Tan, W.-H., Glader, L. J., Barkovich, A. J., Dobyns, W. B., ... Walsh, C. A. (2012). CHMPIA encodes an essential regulator of BMI1-INK4A in cerebellar development. *Nature Genetics*, **44**, 1260–1264. https://doi.org/10.1038/ng.2425

Neyen, C., Runchel, C., Schüpfer, F., Meier, P., & Lemaitre, B. (2016). The regulatory isoform rPGRP-LC induces immune resolution via endosomal degradation of receptors. *Nature Immunology*, **17**, 1150–1158. https://doi.org/10.1038/ni.3536

Ni, J.-Q., Liu, L.-P., Binari, R., Hardy, R., Shim, H.-S., Cavallaro, A., Booker, M., Pfeiffer, B. D., Markstein, M., Wang, H., Villalta, C., Laverty, T. R., Perkins, L. A., & Perrimon, N. (2009). A Drosophila resource of transgenic RNAi lines for neurogenetics. *Genetics*, **182**, 1089–1100. https://doi.org/10.1534/genetics.109.10360

Nickerson, D. P., West, M., Henry, R., & Odorizzi, G. (2010). Regulators of Vps4 ATPase activity at endosomes differentially influence the size and rate of formation of intraluminal vesicles. *Molecular Biology of the Cell*, **21**, 1023–1032. https://doi.org/10.1091/mbc.E09-09-0776

Ou-Yang, Q. - H., Duan, Z. - X., Jin, Z., & Lei, J. - X. (2014). OLC1 is overexpressed in breast cancer and its expression correlates with poor patient survival. *Tumour Tumour Biology*, **35**, 8823–8827. https://doi.org/10.1007/s13277-014-2130-7

Prag, G., Watson, H., Kim, Y. C., Beach, B. M., Ghirlando, R., Hummer, G., Bonifacino, J. S., & Hurley, J. H. (2007). The Vps27/Hsel complex is a GAT domain-based scaffold for ubiquitin-dependent sorting. *Developmental Cell*, **12**, 973–986. https://doi.org/10.1016/j.devcel.2007.04.013

Prince, E., Kroeger, B., Gligorov, D., Wilson, C., Eaton, S., Karch, F., Brankatschk, M., & Maeda, R. K. (2019). Rab-mediated trafficking in the secondary cells of Drosophila male accessory glands and its role in fecundity. *Traffic*, **20**, 137–151. https://doi.org/10.1111/tra.12622

Raiborg, C., Bache, K. G., Mehlum, A., Stang, E., & Stenmark, H. (2001). Hrs recruits clathrin to early endosomes. *Embo Journal*, **20**, 5008–5021. https://doi.org/10.1093/emboj/20.17.5008

Raymond, C. K., Howald-Stevenson, I., Vater, C. A., & Stevens, T. H. (1992). Morphological classification of the yeast vacuolar protein sorting mutants: Evidence for a prevacuolar compartment in class E vps mutants. *Molecular Biology of the Cell*, **3**, 1389–1402.

Razi, M., & Futter, C. E. (2006). Distinct roles for Tsg101 and Hrs in multivesicular body formation and inward vesiculation. *Molecular Biology of the Cell*, **17**, 3469–3483. https://doi.org/10.1091/mbc.E05-11-1054

Redhai, S., Hellberg, J. E., Wainwright, M., Perera, S. W., Castellanos, F., Kroeger, B., Gandy, C., Leiblich, A., Corrigan, L., Hilton, T., Patel, B., Fan, S. J., Hamdy, F., Goberdhan, D. C., & Wilson, C. (2016). Regulation of dense-core granule replenishment by autocrine BMP signalling in *Drosophila* secondary cells. *Plos Genetics*, **12**, e1006366. https://doi.org/10.1371/journal.pgen.1006366

Rideout, E. J., Dornan, A. J., Neville, M. C., Eadie, S., & Goodwin, S. F. (2010). Control of sexual differentiation and behavior by the doublesex gene in Drosophila melanogaster. *Nature Neuroscience*, **13**, 458–466. https://doi.org/10.1038/nn.2515

Rieder, S. E., Banta, L. M., Köhrer, K., McCaffery, J. M., & Emr, S. D. (1996). Multilamellar endosome-like compartment accumulates in the yeast vps28 vacuolar protein sorting mutant. *Molecular Biology of the Cell*, **7**, 985–999. https://doi.org/10.1091/mbc.7.6.985

Rue, S. M., Mattei, S., Saksena, S., & Emr, S. D. (2007). Novel Ist1-Did2 complex functions at a late step in multivesicular body sorting. *Molecular Biology of the Cell*, **19**, 475–484. https://doi.org/10.1091/mbc.e07-07-0694

Rusten, T. E., Vaccari, T., Lindmo, K., Rodahl, L. M. W., Nezis, I. P., Sem-Jacobsen, C., Wendler, F., Vincent, J.-P., Brech, A., Bilder, D., & Stenmark, H. (2007). ESCRTs and Fab1 regulate distinct steps of autophagy. *Current Biology*, **17**, 1817–1825. https://doi.org/10.1016/j.cub.2007.09.032

Saber, S. H., Ali, H. E. A., Gaballa, R., Gaballah, M., Ali, H. I., Zerfaoui, M., & Abd Elmageed, Z. Y. (2020). Exosomes are the driving force in preparing the soil for the metastatic seeds: Lessons from the prostate cancer. *Cells*, **9**, 564. https://doi.org/10.3390/cells9030564

Saksena, S., & Emr, S. D. (2009). ESCRTs and human disease. *Biochemical Society Transactions*, **37**, 167–172. https://doi.org/10.1042/BST0370167

Schindelin, J., Rueden, C. T., Hiner, M. C., & Eliceiri, K. W. (2015). The ImageJ ecosystem: An open platform for biomedical image analysis: T HE I MAGEJ E COSYSTEM. *Molecular Reproduction and Development*, **82**, 518–529. https://doi.org/10.1002/mrd.22489

Schuh, A. L., & Audhya, A. (2014). The ESCRT machinery: From the plasma membrane to endosomes and back again. *Critical Reviews in Biochemistry and Molecular Biology*, **49**, 242–261. https://doi.org/10.3109/10409238.2014.881777

Sheng, Z., Yu, L., Zhang, T., Pei, X., Li, X., Zhang, Z., & Du, W. (2016). ESCRT-0 complex modulates Rbf-mutant cell survival by regulating Rhomboid endosomal trafficking and EGFR signaling. *Journal of Cell Science*, **129**, 2075–2084. https://doi.org/10.1242/jcs.182261

Shiflett, S. L., Ward, D. M., Huynh, D., Vaughn, M. B., Simmons, J. C., & Kaplan, J. (2004). Characterization of Vtalp, a Class E Vps protein in *Saccharomyces cerevisiae*. *Journal of Biological Chemistry*, **279**, 10982–10990. https://doi.org/10.1074/jbc.M312669200

Shim, J. - H., Xiao, C., Hayden, M. S., Lee, K. - Y., Trombetta, E. S., Pypaert, M., Nara, A., Yoshimori, T., Wilm, B., Erdjument-Bromage, H., Tempst, P., Hogan, B. L. M., Mellman, I., & Ghosh, S. (2006). CHMP5 is essential for late endosome function and down-regulation of receptor signaling during mouse embryogenesis. *Journal of Cell Biology*, **172**, 1045–1056. https://doi.org/10.1083/jcb.200509041

Stuffers, S., Wegner, C. S., Stenmark, H., & Brech, A. (2009). Multivesicular endosome biogenesis in the absence of ESCRTs. *Traffic*, **10**, 925–937. https://doi.org/10.1111/j.1600-0854.2009.00920.x

Teis, D., Saksena, S., Judson, B. L., & Emr, S. D. (2010). ESCRT-II coordinates the assembly of ESCRT-III filaments for cargo sorting and multivesicular body vesicle formation. *Embo Journal*, **29**, 871–883. https://doi.org/10.1038/emboj.2009.408

Théry, C., Witwer, K. W., Aikawa, E., Alcaraz, M. J., Anderson, J. D., Andriantsitohaina, R., Antoniou, A., Arab, T., Archer, F., Atkin-Smith, G. K., Ayre, D. C., Bach, J.-M., Bachurski, D., Baharvand, H., Balaj, L., Baldacchino, S., Bauer, N. N., Baxter, A. A., Bebawy, M., ... Zuba-Surma, E. K. (2018). Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. *Journal of Extracellular Vesicles*, **7**, 1535750. https://doi.org/10.1080/20013078.2018.1535750
Thompson, A. G., Gray, E., Heman-Ackah, S. M., Mäger, I., Talbot, K., Andaloussi, S. E., Wood, M. J., & Turner, M. R. (2016). Extracellular vesicles in neurodegenerative disease—Pathogenesis to biomarkers. *Nature Reviews Neurology*, **12**, 346–357. https://doi.org/10.1038/nrneurol.2016.68

Tomas, A., & Futter, C. E. (2015). Stress reveals new destination for EGF receptor. *Cell Cycle*, **14**, 3343–3344. https://doi.org/10.1080/15384101.2015.1093432

Trajkovic, K., Hsu, C., Chiantia, S., Rajendran, L., Wenzel, D., Wieland, F., Schwille, P., Brügger, B., & Simons, M. (2008). Ceramide triggers budding of exosome vesicles into multivesicular endosomes. *Science*, **319**, 1244–1247. https://doi.org/10.1126/science.1153124

Tsang, H. T. H., Connell, J. W., Brown, S. E., Thompson, A., Reid, E., & Sanderson, C. M. (2006). A systematic analysis of human CHMP protein interactions: Additional MIT domain-containing proteins bind to multiple components of the human ESCRT III complex. *Genomics*, **88**, 333–346. https://doi.org/10.1016/j.ygeno.2006.04.003

Tsuneizumi, K., Nakayama, T., Kamoshida, Y., Kornberg, T. B., Christian, J. L., & Tabata, T. (1997). Daughters against dpp modulates dpp organizing activity in *Drosophila* wing development. *Nature*, **389**, 627–631. https://doi.org/10.1038/39362

Vaccari, T., Rusten, T. E., Menut, L., Nezis, I. P., Brech, A., Stenmark, H., & Bilder, D. (2009). Comparative analysis of ESCRT-I, ESCRT-II and ESCRT-III function in Drosophila by efficient isolation of ESCRT mutants. *Journal of Cell Science*, **122**, 2413–2423. https://doi.org/10.1242/jcs.046391

Vietri, M., Radulovic, M., & Stenmark, H. (2019). The many functions of ESCRTs. *Nature Reviews Molecular Cell Biology*, **21**, 25–42. https://doi.org/10.1038/s41580-019-0177-4

Villarroya-Beltri, C., Gutiérrez-Vázquez, C., Sánchez-Cabo, F., Pérez-Hernández, D., Vázquez, J., Martin-Cofreces, N., Martinez-Herrera, D. J., Pascual-Montano, A., Mittelbrunn, M., & Sánchez-Madrid, F. (2013). Sumoylated hnRNPA2B1 controls the sorting of miRNAs into exosomes through binding to specific motifs. *Nature Communications*, **4**, 2980. https://doi.org/10.1038/ncomms3980

Vizcaíno, J. A., Csordas, A., Del-Toro, N., Dianes, J. A., Griss, J., Lavidas, I., Mayer, G., Perez-Riverol, Y., Reisinger, F., Ternent, T., Xu, Q. W., Wang, R., & Hermjakob, H. (2016). 2016 update of the PRIDE database and its related tools. *Nucleic Acids Research*, **44**(D1), D447–D456. https://doi.org/10.1093/nar/gkw880

Ward, D. M., Vaughn, M. B., Shiflett, S. L., White, P. L., Pollock, A. L., Hill, J., Schnegelberger, R., Sundquist, W. I., & Kaplan, J. (2005). The role of LIP5 and CHMP5 in multivesicular body formation and HIV-1 budding in mammalian cells. *Journal of Biological Chemistry*, **280**, 10548–10555. https://doi.org/10.1074/jbc.M413734200

White, I. J., Bailey, L. M., Aghakhani, M. R., Moss, S. E., & Futter, C. E. (2006). EGF stimulates annexin 1-dependent inward vesiculation in a multivesicular endosome subpopulation. *Embo Journal*, **25**, 1–12. https://doi.org/10.1038/sj.emboj.7600759

Xiao, J., Xia, H., Zhou, J., Azmi, I., Davies, B. A., Katzmann, D. J., & Xu, Z. (2008). Structural basis of Vta1 function in the multi-vesicular body sorting pathway. *Developmental Cell*, **14**, 37–49. https://doi.org/10.1016/j.devcel.2007.10.013

Yuan, J., Ma, J., Zheng, H., Shi, T., Sun, W., Zhang, Q., Lin, D., Zhang, K., He, J., Mao, Y., Gao, X., Gao, P., Han, N., Fu, G., Xiao, T., Gao, Y., Ma, D., & Cheng, S. (2008). Overexpression of OLC1, cigarette smoke, and human lung tumorigenesis. *Journal of the National Cancer Institute*, **100**, 1592–1605. https://doi.org/10.1093/jnci/djn379

Zack, G. W., Rogers, W. E., & Latt, S. A. (1977). Automatic measurement of sister chromatid exchange frequency. *Journal of Histochemistry and Cytochemistry*, **25**, 741–753. https://doi.org/10.1177/25.7.70454

## SUPPORTING INFORMATION

Additional supporting information can be found online in the Supporting Information section at the end of this article.

**How to cite this article:** Marie, P. P., Fan, S-. J., Mason, J., Wells, A., Mendes, C. C., Wainwright, S. M., Scott, S., Fischer, R., Harris, A. L., Wilson, C., & Goberdhan, D. C. I. (2023). Accessory ESCRT-III proteins are conserved and selective regulators of Rab11a-exosome formation. *Journal of Extracellular Vesicles*, **12**, e12311. https://doi.org/10.1002/jev2.12311
Thompson, A. G., Gray, E., Heman-Ackah, S. M., Mäger, I., Talbot, K., Andaloussi, S. E., Wood, M. J., & Turner, M. R. (2016). Extracellular vesicles in neurodegenerative disease—Pathogenesis to biomarkers. *Nature Reviews Neurology*, **12**, 346–357. https://doi.org/10.1038/nrneurol.2016.68

Tomas, A., & Futter, C. E. (2015). Stress reveals new destination for EGF receptor. *Cell Cycle*, **14**, 3343–3344. https://doi.org/10.1080/15384101.2015.1093432

Trajkovic, K., Hsu, C., Chiantia, S., Rajendran, L., Wenzel, D., Wieland, F., Schwille, P., Brügger, B., & Simons, M. (2008). Ceramide triggers budding of exosome vesicles into multivesicular endosomes. *Science*, **319**, 1244–1247. https://doi.org/10.1126/science.1153124

Tsang, H. T. H., Connell, J. W., Brown, S. E., Thompson, A., Reid, E., & Sanderson, C. M. (2006). A systematic analysis of human CHMP protein interactions: Additional MIT domain-containing proteins bind to multiple components of the human ESCRT III complex. *Genomics*, **88**, 333–346. https://doi.org/10.1016/j.ygeno.2006.04.003

Tsuneizumi, K., Nakayama, T., Kamoshida, Y., Kornberg, T. B., Christian, J. L., & Tabata, T. (1997). Daughters against dpp modulates dpp organizing activity in *Drosophila* wing development. *Nature*, **389**, 627–631. https://doi.org/10.1038/39362

Vaccari, T., Rusten, T. E., Menut, L., Nezis, I. P., Brech, A., Stenmark, H., & Bilder, D. (2009). Comparative analysis of ESCRT-I, ESCRT-II and ESCRT-III function in Drosophila by efficient isolation of ESCRT mutants. *Journal of Cell Science*, **122**, 2413–2423. https://doi.org/10.1242/jcs.046391

Vietri, M., Radulovic, M., & Stenmark, H. (2019). The many functions of ESCRTs. *Nature Reviews Molecular Cell Biology*, **21**, 25–42. https://doi.org/10.1038/s41580-019-0177-4

Villarroya-Beltri, C., Gutiérrez-Vázquez, C., Sánchez-Cabo, F., Pérez-Hernández, D., Vázquez, J., Martin-Cofreces, N., Martinez-Herrera, D. J., Pascual-Montano, A., Mittelbrunn, M., & Sánchez-Madrid, F. (2013). Sumoylated hnRNPA2B1 controls the sorting of miRNAs into exosomes through binding to specific motifs. *Nature Communications*, **4**, 2980. https://doi.org/10.1038/ncomms3980

Vizcaíno, J. A., Csordas, A., Del-Toro, N., Dianes, J. A., Griss, J., Lavidas, I., Mayer, G., Perez-Riverol, Y., Reisinger, F., Ternent, T., Xu, Q. W., Wang, R., & Hermjakob, H. (2016). 2016 update of the PRIDE database and its related tools. *Nucleic Acids Research*, **44**(D1), D447–D456. https://doi.org/10.1093/nar/gkw880

Ward, D. M., Vaughn, M. B., Shiflett, S. L., White, P. L., Pollock, A. L., Hill, J., Schnegelberger, R., Sundquist, W. I., & Kaplan, J. (2005). The role of LIP5 and CHMP5 in multivesicular body formation and HIV-1 budding in mammalian cells. *Journal of Biological Chemistry*, **280**, 10548–10555. https://doi.org/10.1074/jbc.M413734200

White, I. J., Bailey, L. M., Aghakhani, M. R., Moss, S. E., & Futter, C. E. (2006). EGF stimulates annexin 1-dependent inward vesiculation in a multivesicular endosome subpopulation. *Embo Journal*, **25**, 1–12. https://doi.org/10.1038/sj.emboj.7600759

Xiao, J., Xia, H., Zhou, J., Azmi, I., Davies, B. A., Katzmann, D. J., & Xu, Z. (2008). Structural basis of Vta1 function in the multi-vesicular body sorting pathway. *Developmental Cell*, **14**, 37–49. https://doi.org/10.1016/j.devcel.2007.10.013

Yuan, J., Ma, J., Zheng, H., Shi, T., Sun, W., Zhang, Q., Lin, D., Zhang, K., He, J., Mao, Y., Gao, X., Gao, P., Han, N., Fu, G., Xiao, T., Gao, Y., Ma, D., & Cheng, S. (2008). Overexpression of OLC1, cigarette smoke, and human lung tumorigenesis. *Journal of the National Cancer Institute*, **100**, 1592–1605. https://doi.org/10.1093/jnci/djn379

Zack, G. W., Rogers, W. E., & Latt, S. A. (1977). Automatic measurement of sister chromatid exchange frequency. *Journal of Histochemistry and Cytochemistry*, **25**, 741–753. https://doi.org/10.1177/25.7.70454

## SUPPORTING INFORMATION

Additional supporting information can be found online in the Supporting Information section at the end of this article.

**How to cite this article:** Marie, P. P., Fan, S-. J., Mason, J., Wells, A., Mendes, C. C., Wainwright, S. M., Scott, S., Fischer, R., Harris, A. L., Wilson, C., & Goberdhan, D. C. I. (2023). Accessory ESCRT-III proteins are conserved and selective regulators of Rab11a-exosome formation. *Journal of Extracellular Vesicles*, **12**, e12311. https://doi.org/10.1002/jev2.12311