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Figure S2: Expression of the d2EGFP protein (A-O) in transgenic embryos carrying the Wnt/β-catenin reporter construct. (A) Dorsal view of a stage-11 gastrulating embryo when GFP signals first become visible. (B-E) Dynamic and diversified expression of GFP signal in reporter embryos as development proceeds. Particularly strong expression was observed in the developing neural folds (nf) (B), and in the posterior presomitic mesoderm (psm), the migrating cranial neural crest (nc), and the olfactory epithelium (oe) (C-E). GFP is expressed in the DLP mesoderm and the VBI, from which definitive and primitive blood cells arise, respectively (C,E). (F) Transverse section of a stage-32 embryo (dashed line in E) shows high levels of Wnt activity in the VBI (indicated by meso, mesoderm) and also in the overlying ectoderm (ect). The inset shows DAPI staining of the section. (G) Stage-37 embryos show GFP-expressing blood cells leaving the VBI through the vittelline veins and circulating in the posterior cardinal vein (arrowhead) and throughout the whole body. (G’) A closer view of a selected area (G; dashed rectangle) shows GFP-positive blood cells (arrowheads) circulating in the vittelline veins. (H) GFP-expressing and DAPI-stained individual erythrocytes collected from stage-37 embryos and viewed by confocal microscopy. (I-K) Confocal images of transverse sections of an early tadpole. In the inset in panel I, dashed line 1 refers to panels I and J; dashed line 2 refers to panel K. There is strong expression of GFP in the peripheral retina (cmz, I) and in the differentiating lens fibers (lf in I). Wnt activity was also detected in the lung buds (lb in K), pronephric tubules (pt in K), and the dorsal part of the midbrain- hindbrain boundary (mhb in J). (L-N) Views of a late tadpole stage: dorsal (L), ventral (M) and lateral (N). GFP is continuously observed in the developing liver (lv in L), pronephric tubules (pt in N), and the brain (M,N). Note that GFP is expressed at the boundaries between the rhombomeres of the hindbrain (arrowheads in N). (O-Q) GFP expression is visible in various organs about the time of metamorphosis, especially in the mesencephalon (mesc in O), the pancreas (pc in P), and the growth plates of the developing limbs and digits (arrowhead in Q). Insets correspond to bright field images. (R) Detection of Wnt/β-catenin signaling in an E8.5 embryo of the BAT-Gal Wnt-reporter mouse. Strong β-galactosidase activity is visible in the embryo proper (lower left half) and in the extraembryonic mesoderm that forms the blood islands in the yolk sac (arrowhead). No activity is seen in the surrounding visceral endoderm (arrow).
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Analysis of the wnt/beta-catenin reporter activity in transgenic embryos direct GFP visualization, in a Confocal image showing
a dorsolateral view of head and spinal cord of a NF stage 45 embryo: e, eye; fb, forebrain; i.e., inner ear; mb, midbrain; mhb, midbrain-hindbrain boundary; np, nasal pit; ov, otic vesicles; rh, rhombomeres.
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GFP expression in embryo of Wnt report Tg line Xtr.Tg(WntREs:dEGFP)Vlemx
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