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Experiment details for six1

Differential distribution of competence for panplacodal and neural crest induction to non-neural and neural ectoderm.

Differential distribution of competence for panplacodal and neural crest induction to non-neural and neural ectoderm.

Gene Clone Species Stages Anatomy
six1.L laevis NF stage 18 anterior placodal area

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  Fig. 5. Effects of Dlx3 or GATA2 gain or loss of function on non-neural ectodermal markers. (A-G′′) Neural plate stage embryos after unilateral injection (lower half; marked by light blue β-galactosidase or green mycGFP staining) of various constructs as indicated. Reductions (arrows) and broadening or ectopic expression domains (asterisks) in the neural (green) and non-neural (orange) ectoderm compared with the control side (arrowheads) are indicated. Insets depict additional embryos with ectopic expression of Six1 (A), Eya1 (B) and Dlx5 (D) in central neural plate.

Gene Clone Species Stages Anatomy
six1.L laevis NF stage 18 anterior placodal area

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  Fig. 7. Dlx3 promotes different non-neural fates depending on the signaling environment. (A-B′) Keratin (A,A′) and Six1 (B,B′) expression in neural plate stage embryos after unilateral injection (lower half; marked by light-blue β-galactosidase staining) of Dlx3 mRNA and treatment with DMSO (A,B) or the BMP signaling antagonist dorsomorphin (A′,B′). Ectopic Keratin expression in the neural plate is strongly reduced, while ectopic Six1 expression is strongly enhanced by dorsomorphin treatment (inset in B′ shows another example). (C) Effects of signaling agonists or antagonists on ectopic expression of Keratin or Six1 in the neural plate (NP). After Dlx3 mRNA injection, the increase in ectopic Six1 expression by dorsomorphin was blocked by the Wnt agonist azakenpaullone and the FGF antagonist SU5402, whereas co-injection of FGF8 had no significant effect. After GATA2 mRNA injection, dorsomorphin did not increase ectopic Six1 expression, whereas Keratin (not shown) was never ectopically expressed (Fisher’s exact test; *P≤0.05, n.s., not significant). (D-E′′) Transverse sections through neural plate of embryos shown in A (D-D′′) and the inset of B′ (E-E′′). Sections are shown in brightfield (D,E) in the green fluorescent channel, showing mycGFP-positive Dlx3-injected cells (D′,E′); in the red fluorescent channel, showing Sox3 immunostaining (D′,E′); and in overlay (D′′,E′′). DAPI-stained nuclei are shown for orientation in all panels. Ectopic Keratin and Six1 expression is confined to Dlx3-injected cells (asterisks), whereas residual Sox3 staining on injected side is found only in cells that did not receive Dlx3 (arrows). not, notochord.

Gene Clone Species Stages Anatomy
six1.L laevis NF stage 18 anterior placodal area , posterior placodal area

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  Fig. S3. Effects of Dlx3 or GATA2 loss of function after injection of engrailed repressor constructs on neural and non-neural ectodermal markers. Neural plate stage embryos after unilateral injection (lower half; marked by light blue β-galactosidase or green mycGFP staining) of various constructs as indicated. Reductions (arrows), and broadening or ectopic expression domains (asterisks) in the neural (green) and non-neural ectoderm (orange) compared with the control side (arrowheads) are indicated. Double asterisks indicate lateral displacement owing to widening of the neural plate. See text for details.

Gene Clone Species Stages Anatomy
six1.L laevis NF stage 22 somite , cranial placode , anterior placodal area , posterior placodal area

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  Fig. 1 . Induction of ectodermal markers at neural plate boundaries. Ectodermal marker expression in tailbud stage embryos (albino) that had received neural plate grafts (pigmented and/or mycGFP-labeled) into belly ectoderm at stage 13. (A-F′′) Grafts (arrows, outlined) are shown in overviews (A-F) and transverse sections in brightfield (A′-F′), green fluorescent channel (A′-F′), and overlay (A′′-F′′). Sox3 (A-A′′), Sox11 (B-B′′), Zic1 (C-C′′; inset shows higher magnification of expression in graft) and FoxD3 (D-D′′) are maintained or induced (asterisks) in graft. Six1 (F-F′′) is induced in host ectoderm (arrowheads), whereas Sox9 (E-E′′) is induced in the graft (asterisk) and sometimes in host ectoderm (arrowhead).

Gene Clone Species Stages Anatomy
six1.L laevis NF stage 26 stomodeal-hypophyseal primordium , somite , epibranchial placode , cranial placode , anterior placodal area , [+]

  Fig. 6. Dlx3 and GATA2 are required cell-autonomously in the non-neural ectoderm for Six1 induction. (A-F′)Host embryos were co-injected with mycGFP mRNA and either Dlx3 MO (A-C′) or GATA2 MO (D-F′) at the two- to eight-blastomere stage and received a neural plate (NP) graft (pigmented) into their belly ectoderm (B-B′) at stage 13. After host embryos reached tailbud stage, grafts are shown in overview (A,D) and at higher magnifications (B-C′,E-F′) in brightfield (B-F), green fluorescent channel (B′-F′) and an overlay (B′-F′). Boxed areas in B and E are shown in detail in C and F, respectively. Six1 induction in non-neural host ectoderm around the graft (asterisks) is specifically suppressed in cells that received high levels of Dlx3 MO or GATA2 MO (arrows or outlined areas of green cells).