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Fig. 1 Generation and characterisation of sox10:egfp transgenic fish. (A) Restriction maps of the sox10 genomic region. Exons are boxed, sox10 untranslated sequences are indicated in light blue and coding region in dark blue. Lower panel shows a schematic of the construct used to generate transgenic fish. Yellow box shows the polyadenylation signal. Sizes in kb, N=NotI, S=SalI, RV=EcoRV, B=BamHI, Sp=SpeI, E=EcoRI, Nc=NcoI. (B-H) GFP in germline sox10:eGFP transgenic fish is seen in premigratory and migratory NCCs, in time and manner recapitulating the expression of sox10. Premigratory cranial NC at 11 hpf (B; fluorescence (left panel), DIC (middle) and combined (right)) and 17 hpf (C). (D) Cranial NCCs at 15 hpf to show resolution of fine cell morphology. (E) Premigratory (arrowhead) and migrating NCCs in head and on medial pathway in anterior trunk (arrows) at 24 hpf. (F) At 24 hpf, cranial GFP-expressing cells include migrating NC (mc), NC in branchial arches (a), glial precursors surrounding the cranial ganglia (g), and otic epithelium (o). (G) Double immunofluorescence imaging of GFP (green) and neurons (Hu, red) at 48 hpf. There is continued expression in otic epithelium and arches. (H) Tail of 40 hpf embryo to show nascent DRG and migrating NCCs. (I-K) GFP expression persists in peripheral glia, including DRG satellite glia (J), Schwann cells [posterior lateral line (I) and spinal nerves (J,K)], here at 5 dpf. (J) Double immunofluorescence demonstrates complementary expression of GFP (green) and Hu (red) in DRGs. (K) GFP+ Schwann cells are intimately associated with spinal nerve axons (DM-GRASP, red). All are confocal images except (B) and are of live embryos except (G), (J) and (K). In this and all subsequent figures, embryos are shown in lateral view unless stated otherwise. Scale bars: 100 μm in B,C,E; 20 μm in D,F,I,J,K; 50 μm in G,H.