(B, F), anti-GFP antibodies (A, E) and DAPI (C, G). Arrowheads indicate co-localisation between the MACO-1 and the nuclear envelop marker. EMR, Emerin; MANS, Mannosidase. Scale bars represent 1 micron. Found at: doi:10.1371/journal.pgen.1001341.s006 (0.41 MB PDF) Figure S7 C. elegans macoilin mutants have wild-type neuronal cell morphology, axon guidance, and axon polarity (A). The morphology of cell bodies and axons of VD and DD neurons visualized using a punc-47::gfp transgene appears wild type in maco- 1(db9) mutant animals. Scale bar: 20 mm. (B). The mechanosen- sory neurons appear wild type in maco-1(db9) animals. Neurons were visualized using pmec-4::gfp. Scale bar: 20 mm. (C) maco-1(db9) mutants do not have any apparent defect in the morphology of sensory cilia, dendrites, cell bodies and axons of ADL neurons, as visualized using a psrh-220::gfp marker. Scale bar: 10 mm. (D, E) punc-25::SNB-1::GFP is localized along the ventral processes of the DD neurons in N2 wild type and maco-1 L1 larvae. Polarity of DD neurons in maco-1 L1 worms is normal, and MACO-1 is not essential for transport of synaptic vesicles to synapses (see also Figure 6). Scale bar: 20 mm. (F, G) In both npr-1(ad609) and maco- 1(db9); npr-1(ad609) animals SNB-1::GFP is localized along the axonal processes in URX neurons, as predicted by the electron micrograph reconstruction of this neuron. The transgene used is pgcy-36::snb-1-YFP. Scale bar: 10 mm. Found at: doi:10.1371/journal.pgen.1001341.s007 (4.57 MB PDF) Figure S8 Localization of TAX-4-GFP appears wild type in maco-1 mutants. Localization of TAX-4::GFP appears wild-type in maco-1 mutants. npr-1(ad609) (A–B) and maco-1(db9); npr-1(ad609) (C–F) worms expressing pgcy-32::tax-4::GFP from extrachromo- somal arrays. TAX-4 can be observed in URX cell bodies (arrow head) and dendrites (arrows) in both npr-1(ad609) and maco-1(db9); npr-1(ad609) animals (A–D). TAX-4::GFP traffics correctly to PQR dendrites in maco-1(db9); npr-1(ad609) worms (E–F). Scale bars represent 10 microns (A, C, E) and 2 microns (B, D, F). Found at: doi:10.1371/journal.pgen.1001341.s008 (1.04 MB PDF) Figure S9 Loss of maco-1 does not cause obvious defects to localization of GFP-EGL-19. maco-1 mutants do not have altered subcellular localization of GFP::EGL-19. npr-1(ad609) (A, C, E) and maco-1; npr-1(ad609) (B, D, F) worms expressing GFP::EGL- References 1. Almedom RB, Liewald JF, Hernando G, Schultheis C, Rayes D, et al. (2009) An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse. EMBO J 28: 2636–2649. 2. Vogel F, Hartmann E, D G, Rapoport TA (1990) Segregation of the signal sequence receptor protein in the rough endoplasmic reticulum membrane. 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Scale bars represent 20 microns (A, B, C, D, G) and 2 microns (E, F). In panels A – D red indicates fluorescence from the co-injection marker, pgcy-32::mcherry, which directs expression in the URX and AQR head neurons and the PQR tail neuron. Co-localization between red and green fluorescence confirms that EGL-19 is expressed in AQR, PQR and URX neurons. Found at: doi:10.1371/journal.pgen.1001341.s010 (1.97 MB PDF) Figure S10 MACO-1 homologues are present in many eumetazoan lineages. Multiple alignment of retrieved sequences of MACO-1 homologues. Black and gray shadings indicate a minimum of 70% sequence identity and similarity, respectively. Found at: doi:10.1371/journal.pgen.1001341.s009 (0.06 MB PDF) Supporting materials and methods. at: doi:10.1371/journal.pgen.1001341.s011 (0.04 MB Text S1 Found DOC) Acknowledgments We thank Akiko Miyara and Ikue Mori for sharing unpublished data; Melissa Rolls for plasmids; Yuji Kohara for cDNAs; Colin Dolphin for help with recombineering; Emanuel Busch, Eiji Kodama, Patrick Laurent, Zoltan Soltesz, and Kate Weber for critical reading of the manuscript. We gratefully acknowledge receipt of strains from the Caenorhabditis Genetics Center, Mei Zhen, Erik Jorgensen, and Yishi Jin. LBR thanks Dr. Jean- Louis Bessereau for his support. Author Contributions Conceived and designed the experiments: FAC LBR AC MdB. Performed the experiments: FAC LBR AC MdB. Analyzed the data: FAC LBR AC MdB. Contributed reagents/materials/analysis tools: BHHC ML. Wrote the paper: FAC LBR MdB. 13. Kuvbachieva A, Bestel AM, Tissir F, Maloum I, Guimiot F, et al. 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