DEV023119 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Characterization of White Rooster serum. (A) Wild-type (left) and amelanotic (right) Jungle Fowl roosters. (B-H) Blood serum obtained from the depigmented rooster (WRS) contains antibodies that identify MITF-positive melanoblasts in the DL pathway (B) and in the retina (C) of an embryonic day-4 chick. At later stages, WRS identifies cells in the ectoderm of embryonic day-9 chick (D). In tissue culture, WRS colocalizes with MITF (E) and pigmentation (G), but not with the neuronal marker 16A11 (F). By western blot analysis (H), WRS and the Smyth line serum immunolabel the same protein bands in cell lysates from stage-25 quail and day-9 chick. nt, neural tube; rpe, pigmented retinal epithelium

• Supplemental Figure S2 -

  Fig. S1. Characterization of White Rooster serum. (A) Wild-type (left) and amelanotic (right) Jungle Fowl roosters. (B-H) Blood serum obtained from the depigmented rooster (WRS) contains antibodies that identify MITF-positive melanoblasts in the DL pathway (B) and in the retina (C) of an embryonic day-4 chick. At later stages, WRS identifies cells in the ectoderm of embryonic day-9 chick (D). In tissue culture, WRS colocalizes with MITF (E) and pigmentation (G), but not with the neuronal marker 16A11 (F). By western blot analysis (H), WRS and the Smyth line serum immunolabel the same protein bands in cell lysates from stage-25 quail and day-9 chick. nt, neural tube; rpe, pigmented retinal epithelium

• Supplemental Figure S3 -

  Fig. S3. Overexpression of EDNRB2 in melanoblasts leads to increased numbers of cells in more lateral positions. (A) Schematic of a transverse section demarcating the MSA, the ventral (V) pathway and three regions of the DL path: medial DL, lateral DL and lateral mesenchyme. (B-F) Lateral views of stage-21 (B,C) and stage-24 (E,F) chick embryos co-electroporated at stage 16 with pBluescript/siSTRIKE-empty (B,E) or pCAGG-E2⁺/siSTRIKE-empty (C,F). At these stages, the wave of GFP⁺ melanoblasts (arrows) in pCAGG-E2⁺/siSTRIKE-empty embryos (C,F) appears to be more cell-dense and in more lateral positions than in control embryos (B,E). Quantification of WRS⁺ (MITF⁺)/GFP⁺ cells demonstrates an increase overall in the number of melanoblasts present at both of these stages (D,G). Data are presented as mean±s.d., with (*) set at P&λτ;0.05. dm, dermamyotome; drg, dorsal root ganglia; nt, neural tube; ant, anterior.