JCS03205 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. (A,B) TUNEL reaction of neurosphere (arrows in A and B) without BMP-2 pre-treatment 24 hours after transplantation. One TUNEL-reactive chick host nucleus outside the neurosphere is visible in B (arrowhead). Non-reactive nuclei are stained with methylene blue. No specific reaction is present in the neurosphere, indicating vital cells. The presumptive staining at the upper border of the neurosphere in B is an artefact.

• Supplemental Figure 2 -

  Fig. S2. This supplement shows the diversity of patterns of emigrating cells compared with non- emigrating cells. Non-BMP-2 pre-treated neurospheres 24 hours after transplantation into the chick embryo neural tube are compared with BMP-2 pre-treated neurospheres. Images were taken from experimental series as follows: A-D, exp. 1; E,F, exp. 3; G,H, exp. 2; I-L, exp. 1; M,N, exp. 3; O,P, exp. 2 (see Table 1).

• Supplemental Figure 3 -

  Fig. S3. (A) TuJ1 staining of a chick embryo 24 hours after transplantation of a neurosphere without BMP-2 treatment. Differentiating motoneurons were stained in the host neural tube. (B) Higher magnification of region indicated in A. Two TuJ1-positive cells of the neurosphere remaining in the lumen of the neural tube are marked (arrows). (C) TuJ1 staining of an embryo with a BMP-2 treated, integrating neurosphere. (D) Higher magnification as indicated in C. Two TuJ1-positive cells of the neurosphere that are integrating into the roof plate are shown (arrows).

• Supplemental Figure 4 -

  Fig. S4. (A) Whole-mount chick embryo stage 12 (HH) stained with HNK-1. At this stage neurosphere transplantation occurred (2 days of total incubation time). (B) Enlargement of A. At the spot of neurosphere transplantation, which corresponds to the caudally most recently formed somite, no specific HNK-1 pattern is visible dorsal to the neuroepithelium indicating that neural crest emigration has not yet begun at this caudal trunk level (compare with more cranial levels). HNK-1 staining of the respective section of the lower trunk region of an untransplanted chick (insert). (C) Whole-mount chick embryo stage 18 (HH) stained with HNK-1. This stage corresponds to 24 hours after neurosphere transplantation (3 days of total incubation time). (D) Enlargement of C. The spot of neurosphere transplantation corresponds to the site of the upper appendage about to develop. Specific emigration pattern of emigrating neural crest cells is visible throughout the embryo.

• Supplemental Figure 5 -

  Fig. S5. (A) HNK-1 staining of chick embryo stage 12 (HH), 12 hours after transplantation of BMP-2-treated neurosphere. Only neurosphere (NS) cells are HNK-1 positive (arrow). (B) HNK-1 staining with counterstaining of the same region as in A (level of the 16^th pair of somites) of an untransplanted chick embryo stage 12 (HH). No HNK-1-positive cells are present in this stage and region of the embryo. (C) HNK-1 staining of an untransplanted chick embryo stage 18 (HH) at the level of the 16^th pair of somites. HNK-1-positive reaction marks neural crest cells and spinal ganglia. (D) Higher magnification of the region indicated in C. HNK-1-positive neural crest cells can be distinguished along the dorsal edge of the neural epithelium and in the spinal ganglion.