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Supplemental Fig S1. Expression of hhex, sdc4 and fzd7in Xenopus embryo In situ hybridization showing the expression pattern of hhex, sdc4 and fzd7 at late gastrula stage, 0–1 somite stage and 4–6 somite stage. |
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sdc4 (syndecan 4) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 19, mid-sagittal section, anterior right, dorsal up. |
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Fig. 1. Sdc4 is required for foregut organogenesis (A) Schematic of a NF19 stage Xenopus embryo in mid-sagittal section with the foregut (fg) endoderm shown in dark grey and mid/hindgut endoderm in light grey. (B) In situ hybridization showing sdc4 expression at stage NF19. (C) Confocal image showing Alexa lineage tracer (red) in a bisected stage NF19 embryo confirming the targeted microinjection of the foregut endoderm. (D) Western blot analysis of NF19 dissected foreguts confirming Sdc4 depletion by Sdc4-MO injection. (E–G) Lateral view of stage 45 embryos showing foregut hypoplasia in Sdc4-depleted embryos that is rescued by sdc4 mRNA injection. (H–J) In situ hybridization of a2m in isolated stage 45 gut tube showing hypoplastic liver (lv), stomach (st) and pancreas (p). (K–P) In situ hybridization with liver (nr1h5) and pancreas/duodenum (pdx1) markers at stage NF35 showing defects in organ specification, which are rescued by sdc4 RNA injection. In all panels, numbers indicate the embryos exhibiting the phenotype. |
