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Experiment details for shh

Evolutionarily conserved Tbx5-Wnt2/2b pathway orchestrates cardiopulmonary development.

Evolutionarily conserved Tbx5-Wnt2/2b pathway orchestrates cardiopulmonary development.

Gene Clone Species Stages Anatomy
shh.L laevis NF stage 10.5 endomesoderm , anterior

  Fig. S5. Utilizing Xenopus system to understand the genetic interaction between Tbx5 and Shh signaling. A. To study the interaction of Tbx5 and Hedgehog signaling, we utilized: (1) a dexamethasone (DEX) inducible glucocorticoid receptor-Tbx5 (GR-Tbx5) fusion protein, and (2) pharmacological agonists (purmorphamine) and antagonists (cyclopamine) of SMO to activate or repress Hh signaling similar to Fig. 4 (top). Experimental details for testing Hedgehog response in anterior mesendoderm (AME, red, Shh+) and posterior mesendoderm (PME, blue, Shh-) tissues measured by gli1 expression (bottom). Dissected AME or PME tissue was cultured for 6 hours in combinations of dexamethasone (DEX, translocates GR-Tbx5 fusion protein to the nucleus), purmorphamine (Hh agonist), and cyclopamine (Hh antagonist). B. Expression of gli1 examined by qRT-PCR in culture conditions as indicated by the checked boxes. All conditions were normalized to the AME controls (far left). C. Experimental plan for investigating markers of lung development in embryos injected with mistmatched morpholinos (MM-MOs) or Tbx5-MOs. Embryos were cultured in DMSO (grey) or Hh agonist (purmorphamine, blue) for 48 hours. D. qRT-PCR of shh, dhh, gli1, and nkx2-1 from Tbx5-morphant or control embryonic explants treated with DMSO or Hh agonist.

Gene Clone Species Stages Anatomy
shh.L laevis NF stage 35 and 36 spinal cord , notochord , endoderm , floor plate , gall bladder primordium , [+]

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  Fig. S1. Xenopus whole mount in situ hybridization. RNA in situ hybridization was performed in NF35 Xenopus embryos for tbx5, wnt2b, nkx2-1, and shh (whole mount side-view on top, transverse cross-section on bottom). lpm = lateral plate mesoderm, thy = thyroid, nc = notochord, phe = pharyngeal endoderm, fge = foregut endoderm, gb = gall bladder, fp = floor plate.

Gene Clone Species Stages Anatomy
shh.L laevis NF stage 35 and 36 spinal cord , notochord , endoderm , foregut , floor plate , [+]

  shh (sonic hedgehog) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 35 & 36, lateral view, anterior left, dorsal up (top image) and transverse section (below). Key: nc = notochord, phe = pharyngeal endoderm, fge = foregut endoderm, gb = gall bladder, fp = floor plate.