DEV02087 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Endothelial precursor migration in TG(flk1:EGFP)^s843 embryos. Epifluorescence micrographs of Tg(flk1:EGFP)^s843 embryos are shown in dorsal views (A-D,I,J) and lateral views (E-H,K,L). Embryo at 14 hpf (A,E), 16 hpf (B,F), 17 hpf (C,G), 18 hpf (D,H), 20 hpf (I,K) and 24 hpf (J,L).

• Supplemental Figure 2 -

  Fig. S2. Pan-endothelial marker VE-cadherin (cdh5) expression during angioblast migration. Micrographs of in situ hybridization with VE-cadherin. Embryos at 14 hpf (A,D), 16 hpf (B,E) and 18 hpf (C,F), in dorsal (A-C) and lateral views (D-F).

• Supplemental Figure 3 -

  Fig. S3. Pharyngeal expression of flk1:GFP in cloche and bonnie and clyde mutant embryos. Epifluorescence micrographs of a Tg(flk1:EGFP)^s843;clo mutant embryo (B) and a sibling (A) at 36 hpf, and a Tg(flk1:EGFP)^s843;bon mutant embryo at 48 hpf (C). The outlined area in C is shown in F. Longitudinal section of a clo mutant embryo at 60 hpf, visualized for GFP (green) and filamentous actin (red) (D). The outlined area in D is shown in E. White bracket in F marks the pharyngeal region. The pharyngeal GFP expression is present in Tg(flk1:EGFP)^s843;clo mutant embryos (arrowhead in B; D,E), while severely reduced in Tg(flk1:EGFP)^s843;bon mutant embryos (F), suggesting that it is localized in endodermal tissues.

• Supplemental Figure 4 -

  Fig. S4. Adherens junction formation during vascular development. Transverse sections of wild-type embryos (A,B,E,F,I, J) and endoderm-less embryos [Tg(flk1:EGFP)^s843;cas mutants] (C,D,G,H,K,L) visualized for b-catenin (red) (A,C,E,G,I,K) and GFP (green) (B,D,F,H,J,L). The sections are at the level of the 6th (A-H) and 12th somite (I-L). White arrowheads indicate adherens junctions between endothelial cells, which appear after the cells reach the midline.

• Supplemental Figure 5 -

  Fig. S5. Expression of arterial and venous specific markers at the time of differentiation. Micrographs of in situ hybridization with flt4 (A,H), ephrinB2a (B,C,G) and VE-cadherin (cdh5) (D-F). Embryos at 13 hpf (A,D), 15 hpf (B,E) and 17 hpf (C,F), in lateral (A-B) and dorsal views (C-H; flat-mounted in D-H). Arrowheads in C,G,H indicate vascular expression of ephrinB2a (C,G), and flt4 (H). Both ephrinB2a and flt4 are expressed in the angioblasts located at the midline, but not those in the lateral plate mesoderm. Detailed expression patterns of arterial and venous endothelial cell markers can be found at the ZFIN website (www.zfin.org).