Fig. S1

Characterization of the Transgenic Fish Lines tg(sdf1a:sdf1a-GFP) and tg(cxcr4b:cxcr4b-Kate2-IRES-GFP-CaaX), Related to Figures 2, 3, and 4

(A) Schematic overview of the sdf1a:sdf1a-GFP transgene. (B) 40 hpf wild-type embryo stained for sdf1a mRNA. (C, D) 40 hpf tg(sdf1a:sdf1a-GFP)p10; sdf1a/ embryos stained for GFP mRNA (C) or Sdf1a-GFP protein (D). (E–G) trop2 mRNA staining labeling the primordium and deposited neuromasts in 40 hpf wild-type (E), sdf1a/ (F) and tg(sdf1a:sdf1a-GFP)p10; sdf1a/ embryos (G). (H) Schematic overview of the cxcr4b:cxcr4b-Kate2-IRES-eGFP-CaaX transgene. (I–K) (I) 40 hpf wild-type embryo stained for cxcr4b mRNA. (J, K) 40 hpf tg(cxcr4b:cxcr4b-Kate2-IRES-GFP-CaaX)p7; cxcr4b/ embryos stained for kate2 mRNA (J) or imaged live for Kate2 and GFP fluorescence (K). Arrows in J to K indicate the primordium. (L–N) trop2 mRNA staining labeling the primordium and deposited neuromasts in 40 hpf wild-type (L), cxcr4b/ (M) and tg(cxcr4b:cxcr4b-Kate2-IRES-GFP-CaaX)p7; cxcr4b/ embryos (N). Arrows in (E–G) and (L–N) indicate the extent of primordium migration. (O and P) Primordium from a 28 hpf tg(sdf1a:sdf1a-GFP)p10; tg(cxcr4b:cxcr4b-Kate2-IRES-GFP-CaaX); cxcr7b/ embryo (O) and primordium from a 28 hpf tg(cxcr4b:cxcr4b-Kate2-IRES-GFP-CaaX); cxcr7b/ embryo (P) stained for Kate2 (red) and GFP (green). (O) and (P) represent cells bounded by the white box in the overview image of the primordium. Arrowheads indicate the colocalization of Sdf1a-GFP and Cxcr4b-Kate2 puncta in O and Cxcr4b-Kate2 in P. (Q–R) High resolution images of the primordium in a 40 hpf wild-type embryo stained for cxcr4b mRNA (Q) and 40 hpf tg(cxcr4b:cxcr4b-Kate2-IRES-GFP-CaaX)p7; cxcr4b/ embryo stained for kate2 mRNA (R). (S) Mean FmemRed/FmemGreen ratio values across primordia from wild-type (solid circles), sdf1a/ (triangles) and tg(hsp70:sdf1a) embryos (open circles). The front of the primordium is at 0 μm. The gray bars indicate SEM.