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Fig. 10. Functional epistasis of Wnt/b-catenin, FGF, and BMP pathways. A–O: Wnt/b-catenin is necessary and sufficient downstream of FGF to induce respiratory fate. Embryos were injected at the 4-cell stage in both dorsal anterior blastomeres with control-MOs or wnt2/2b-MOs (10 ng/ blastomere) and then injected at the 8-cell stage dorsal-vegetally with 800 pg iFGFR1 mRNA (400 pg/blastomere). Subsequently embryos were treated from stages NF25–NF35 with either, DMSO, PD173074 (100 mM), DMH1 (20 mM), or AP20187 (2 mM to induce the iFGFR1 construct activ- ity). Embryos were analyzed at NF34/35 by in situ hybridization for nkx2.1 expression (A, D, G, J, and M) or by for Nkx2.1 (red), Sox2 (blue), and FoxF1 (green) (B, E, H, K, N). At NF42 siblings were analyzed by in situ hybridization for sftpc expression (C, F, I, L, O). P–Y: The expansion of respiratory fate by Wnt/b-catenin requires BMP signaling. Embryos were injected at the 4-cell stage in both dorsal-anterior blastomeres with control-MOs or wnt2/2b-MOs (10 ng/blastomere) and then injected at the 8-cell stage dorsal-vegetally with 800 pg GR-LefdN-BCTA mRNA (400 pg/blastomere). Subsequently the embryos were treated with DMSO, DMH-1, or PD173047 from NF25-35, and then with dex from NF28-35 and analyzed by in situ hybridization for nkx2.1 at NF35 (P,R,T,V,X) and sftpc at NF42 (Q,S,U,W,Y). Image published in: Rankin SA et al. (2015) Copyright © 2015. Image reproduced with permission of the Publisher, John Wiley & Sons. Gene Synonyms Species Stage(s) Tissue nkx2-1.L nkx2.1, titf1, ttf1, Xnkx-2.1, xnkx2-1 X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 endoderm foregut lung primordium ventral eye forebrain telencephalon diencephalon thyroid primordium olfactory bulb sftpc.S SP-C, SPC, surfc, xSP-C X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 endoderm foregut lung primordium Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + wnt2 MO + NF33/34-35 and 36 (in situ hybridization) fig.10.d Anatomical Phenotype abnormal development of lung Expression Phenotype decreased amount nkx2-1.L expression in lung bud Xla Wt + wnt2 MO + NF42 (immunohistochemistry) fig.10.e Expression Phenotype decreased amount nkx2-1.L expression in foregut mislocalised sox2.S expression in foregut endoderm Xla Wt + wnt2 MO + NF42 (in situ hybridization) fig.10.f Expression Phenotype decreased amount sftpc.L expression in lung bud Xla Wt + PD173074 + NF33/34-35 and 36 (in situ hybridization) fig.10.g Anatomical Phenotype abnormal development of lung Expression Phenotype decreased amount nkx2-1.L expression in lung bud Xla Wt + PD173074 + NF42 (immunohistochemistry) fig.10.h Expression Phenotype decreased amount nkx2-1.L expression in foregut mislocalised sox2.S expression in foregut endoderm Xla Wt + PD173074 + NF42 (in situ hybridization) fig.10.i Expression Phenotype decreased amount sftpc.L expression in lung bud Xla Wt + DMH-1 + NF33/34-35 and 36 (in situ hybridization) fig.10.j Anatomical Phenotype abnormal BMP signaling pathway abnormal development of lung Expression Phenotype decreased amount nkx2-1.L expression in lung bud Xla Wt + DMH-1 + NF42 (immunohistochemistry) fig.10.k Expression Phenotype decreased amount nkx2-1.L expression in foregut mislocalised sox2.S expression in foregut endoderm Xla Wt + DMH-1 + NF42 (in situ hybridization) fig.10.l Expression Phenotype decreased amount sftpc.L expression in lung bud Xla Wt + Mmu.fgfr1del-Mmu.fkbp1adel + wnt2 MO + B/B Homodimerizer + NF33/34-35 and 36 (in situ hybridization) fig.10.m Expression Phenotype decreased amount nkx2-1.L expression in lung bud Xla Wt + Mmu.fgfr1del-Mmu.fkbp1adel + wnt2 MO + B/B Homodimerizer + NF42 (immunohistochemistry) fig.10.n Expression Phenotype decreased amount nkx2-1.L expression in foregut mislocalised sox2.S expression in foregut endoderm Xla Wt + Mmu.fgfr1del-Mmu.fkbp1adel + wnt2 MO + B/B Homodimerizer + NF42 (in situ hybridization) fig.10.o Expression Phenotype decreased amount sftpc.L expression in lung bud Xla Wt + Mmu.lef1-Mmu.ctnnb1-GR + DEX + NF35/36 (in situ hybridization) fig.10.r Anatomical Phenotype abnormal development of lung abnormal Wnt signaling pathway abnormally localised lung bud increased size of the lung bud Expression Phenotype mislocalised nkx2-1.L expression in lung bud increased amount nkx2-1.L expression in lung bud Xla Wt + Mmu.lef1-Mmu.ctnnb1-GR + DEX + NF42 (in situ hybridization) fig.10.s Expression Phenotype lung bud that expresses sftpc.L lung bud that expresses sftpc.L increased amount sftpc.L expression in lung bud Xla Wt + Mmu.lef1-Mmu.ctnnb1-GR + wnt2 MO + DEX + NF35/36 (in situ hybridization) fig.10.t Anatomical Phenotype abnormal development of lung Expression Phenotype lung bud that expresses nkx2-1.L lung bud that expresses nkx2-1.L increased amount nkx2-1.L expression in lung bud Xla Wt + Mmu.lef1-Mmu.ctnnb1-GR + wnt2 MO + DEX + NF42 (in situ hybridization) fig.10.u Expression Phenotype lung bud that expresses sftpc.L lung bud that expresses sftpc.L increased amount sftpc.L expression in lung bud Xla Wt + Mmu.lef1-Mmu.ctnnb1-GR + DEX + PD173074 + NF35/36 (in situ hybridization) fig.10.v Anatomical Phenotype abnormal development of lung Expression Phenotype lung bud that expresses nkx2-1.L lung bud that expresses nkx2-1.L mislocalised nkx2-1.L expression in lung bud increased amount nkx2-1.L expression in lung bud Xla Wt + Mmu.lef1-Mmu.ctnnb1-GR + DEX + PD173074 + NF42 (in situ hybridization) fig.10.w Expression Phenotype lung bud that expresses sftpc.L lung bud that expresses sftpc.L increased amount sftpc.L expression in lung bud Larger Image Printer Friendly View

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