FIGURE 2:

cil-7 is expressed in the 27 EV-releasing ciliated sensory neurons and is targeted to EVs in a myristoylation motif–dependent manner. (A) Pcil-7::GFP was expressed in CEM, RnB, HOB, and IL2 neurons. (B) Pcil-7::CIL-7::GFP in cil-7(tm5848) males localized throughout neurons, including cilia, but was excluded from the nucleus. Pcil-7::CIL-7::GFP rescues cil-7(tm5848) defects (Figure 1K), indicating that this reporter is functional. (C) Pcil-7::CIL-7(G2D)::GFP in cil-7(tm5848) males localized throughout neurons, including cilia, but was excluded from the nucleus. Pcil-7::CIL-7(G2D)::GFP does not rescue cil-7(tm5848) defects (Figure 1K), indicating that this reporter is not functional. (D) CIL-7::GFP as EV cargo was trapped in the molted tail cuticle of the late L4 male in a range from either 1–9 EV particles or >10 EV particles per male tail. The CIL-7(G2D)::GFP myristoylation mutant is not efficiently targeted to EVs. (E) In 100% of animals, CIL-7::GFP was observed in the molted tail cuticle in varying ranges; depicted here is an example of >10 EV particles. (F) In <50% of animals, CIL-7 (G2D)::GFP was observed as EV cargo. In E, ***p < 0.0001 by the Mann–Whitney test. Yellow arrow, cilia; red arrow, dendrite; white arrow, cell body; orange arrowhead, axon; white arrowhead, EV particle.