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Fig. 2. Dril1 in situ hybridization. Expression is high in the nonneural ectoderm of the early neurula, but it is excluded from neural plate; boundary marked with an arrow (A). Late tailbud embryo stained throughout the epidermis (B). Sense controls (C). Sagittal bisections of NF10 (D), NF11 (E), and NF12.5 (F) gastrulae, showing mesodermal expression (black arrowheads). Arrows mark the dorsal lip of the blastopore. Transverse cut of neurula (G) showing staining in presomitic mesoderm (white arrow).
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Fig. 3. Effects of dril1 overexpression and depletion on X. laevis development. Uninjected controls (A); tadpoles exhibiting eye defects (white arrowheads) but unperturbed axes after injection with 2 ng dril1 mRNA (B) or 2 ng VP16–dril (C). Gene Tools control morpholino (D and H), 30 ng dril1 morpholino (E and I), 200 pg EnR–dril RNA (F), 600 pg EnR–dril RNA (G), and 30 ng 5 bp mismatch dril1 morpholino (J).
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Fig. 4. Dril1 morpholino specificity. Autoradiogram of in vitro transcription/translation products derived from CS2–dril1, which contains the morpholino target site (lanes 1–4), or CS2-δ5′dril-MT, which lacks the morpholino target site (lanes 5–8), plus the following combinations of morpholino: (1 and 5) 160 ng 5 bp mismatch morpholino (5-mis MO); (2 and 6) 160 ng dril1 MO; (3 and 7) 800 ng 5-mis MO; and (4 and 8) 800 ng dril1 MO. Lane 9 contains reticulocyte lysate without vector or morpholino. Comparisons cannot be made between lanes 1–4 and 5–8 because products are derived from different templates. The δ5′dril-MT product is of a higher mass than dril1 protein because of the addition of the 6-myc tag.
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Fig. 5. Effects of dril1 morpholino on X. tropicalis development. NF21 embryos injected with 20 ng of a control morpholino, anterior to left (A), or dril1 morpholino (B). The latter embryos failed to complete gastrulation. Uninjected tadpoles (C), tadpoles injected with 3 ng dril1 morpholino (D), morphants rescued by coinjection with 300 pg δ5′dril-MT RNA (E). Rescue of a more severe morphant phenotype (F–H). Uninjected embryos (F), embryos injected with 6 ng dril1 morpholino (G), embryos rescued with 260 pg δ5′dril-MT RNA (H); note elongated anteroposterior axes, and more developed head structures, as evidenced by presence of cement gland. White arrowheads in G mark epidermal lesions.
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Fig. 6. Time lapse of gastrulation in dril1 morphants. Embryos were injected with 30 ng control morpholino (A–C) or dril1 morpholino (D–F) and photographed at NF10 (A and D), NF11 (B and E), and NF12.5 (C and F).
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Fig. 7. In situ hybridization of dril1 morphants. Embryos were injected with either control morpholino (A–F) or dril1 morpholino (G–L). Whole NF13 embryos injected with 20 ng morpholino were stained for chordin (A and G) and Xbra (B and H). Sagittally bisected embryos injected with 30 ng morpholino were stained for frzb-1 (C and I), Fz7 (D and J), and prickle (E and K) at NF13, and for sprouty2 at NF12 (F and L). Symbols in C and I represent archenteron (a), blastocoel (b), dorsal lip of blastopore (white arrowhead), and ventral lip of blastopore (dark arrowhead).
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Fig. 10. Dril1 depletion inhibits the response of animal caps to activin. Control caps minus activin (A and B). Control caps plus activin (C and D). Activin-treated caps from dril1 morphants (E) and EnR–dril-injected embryos (F). Gene induction in activin-treated caps injected with 30 ng dril1 morpholino (G) or 600 pg EnR–dril (H).
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Fig. 12. Dril1 morpholino inhibits smad2-mediated secondary axis formation. (A) Representative secondary axes induced by injection of 500 pg smad2 into a ventral vegetal blastomere at the 8-cell stage. (B) Inhibition of smad2-induced secondary axes by coinjection of 15 ng dril1 morpholino.
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arid3a (AT rich interactive domain 3A (BRIGHT-like)) gene expression in Xenopus laevis, assayed by in situ hybridization, NF stage 10 embryo, mid-sagital section,
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arid3a (AT rich interactive domain 3A (BRIGHT-like)) gene expression in Xenopus laevis, assayed by in situ hybridization, NF stage 11 embryo, mid-sagittal section,
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arid3a (AT rich interactive domain 3A (BRIGHT-like)) gene expression in Xenopus laevis, assayed by in situ hybridization, NF stage 12.5 embryo, mid-sagital section,
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arid3a (AT rich interactive domain 3A (BRIGHT-like)) gene expression in a Xenopus laevis embryo, assayed by in situ hybridization, NF stage 13, anterior view, dorsal up (left).
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arid3a (AT rich interactive domain 3A (BRIGHT-like)) gene expression in Xenopus laevis, assayed by in situ hybridization, NF stage 19 embryo, mid-sagittal section, dorsal up.
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arid3a (AT rich interactive domain 3A (BRIGHT-like)) gene expression in Xenopus laevis, assayed by in situ hybridization, NF stage 31 embryo, lateral view, anterior left, dorsal up.
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Supplementary Fig. 1. Cell division is not inhibited in dril1 morphants. Uninjected embryos (A) or embryos injected with 30 ng dril1 morpholino (B) were stained with the αPH3 antibody at NF10.25.
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