DEV02215 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. lsn mutants have a reduced number of serotonergic ENS neurons. (A-D) Lateral views of the gut tube of 96 hpf wild type (A,C) and lsn (B,D) embryos that have been double stained with anti-Hu antibody (A,B) and anti-5HT (C,D), showing that there is a reduction in both overall neuron number and serotonergic neurons in lsn mutants when compared with wild type. Arrowheads indicate double-labelled cells.

• Supplemental Figure 2 -

  Fig. S2. lsn mutants have no cranial ganglia defects. (A,B) Lateral view of Hu expression in cranial ganglia of wild-type (A) and lsn (B) embryos at 96 hpf. (C,D) Lateral flat mounts of embryos showing the cranial ganglia expression phox2b at 48 hpf. ad, anterodorsal lateral line ganglion; av, anteroventral lateral line ganglion; e, enteric neurons; f, facial ganglion; g, glossopharyngeal ganglion; m, middle lateral line ganglion; o, octaval/statoacoustic ganglion; p, posterior lateral line ganglion; v, vagal ganglia.

• Supplemental Figure 3 -

  Fig. S3. lsn mutants have no defect in initial vagal crest specification and migration or hind-brain patterning. (A,B) Lateral view of crestin expression in wild-type (A) and lsn (B) embryos at 20 hpf. (C,D) Dorsal view of epha4 and hoxb4 expression in wild-type (C) and lsn (D) embryos at 20 hpf. Arrows in A,B indicate vagal crest.

• Supplemental Figure 4 -

  Fig. S4. lsn mutants have no defect in the migration and initial differentiation of pharyngeal cartilage neural crest or in initial specification of the thymus. (A,B) Lateral view of dlx2a expression in the migrating neural crest of wild-type (A) and lsn (B) embryos at 36 hpf. (C,D) Ventral views of pax9 expression in the differentiating pharyngeal arch neural crest of wild-type (C) and lsn (D) embryos at 48 hpf. (E,F) Lateral view of foxn1 expression in the thymic primordium of wild-type (E) and lsn (F) embryos at 48 hpf. ot, otic vesicle; b, branchial arches; h, hyoid arch; m, mandibular arch. Arrows in E,F indicate foxn1 expression.

• Supplemental Figure 5 -

  Fig. S5. lsn mutants have normal intestinal gdnf expression and have no increase in cell death in the intestine (A,B) Lateral view of gdnf expression in the mesendoderm of wild-type (A) and lsn (B) embryos at 48 hpf. (C,D) Ventral views of the intestines of TUNEL stained wild-type (C) and lsn (D) embryos at 48 hpf. (E,F) Lateral views of the intestines TUNEL stained wild-type (E) and lsn (F) embryos at 72 hpf.