Constructs used to dissect Wld^S functional domains and localization in Drosophila neurons. (A) All constructs were generated from mouse Wld^S, which contains N70, the first 16 amino acids (N16) that encode the VCP-binding site, W18, an 18–amino acid linker, and full-length Nmnat1. Nmnat1^dead harbors an H24A mutation; Wld^SΔN16 lacks the N16 domain; N16-Nmnat1 is N16 tethered to the N terminus of Nmnat1; Wld^S-dead is full-length Wld^S with an H24A mutation in Nmnat1. See Materials and methods for details. (B–G) UAS-Wld^S–Myc (B and C), UAS-Wld^S-dead–Myc (D and E), and UAS-Nmnat1–myc (F and G) were expressed with UAS-mCD8–GFP in projection neurons using GH146-Gal4. The arrows point to nuclei, and the arrowheads point to axons. Bar, 12.14 µm.