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Figure 5. FAM and Ecto Operate in the Same Pathway Regulating Smad4(A–L) Panels show in situ hybridizations on Xenopus embryos for the pan-mesodermal marker Xbra (at gastrula stage), for the ventral marker Sizzled and for the dorsoanterior marker Otx2 (at neurula stages). Embryos were microinjected either with morpholino antisense oligonucleotides (MOs), dominant-negative Smad4 mRNA (DN-Smad4), or Ecto mRNA.(M–R) Close-up views of Drosophila wings showing anterior (acv) and posterior (pcv) cross-veins.(N) Mutants for gbb, a BMP ligand, display missing cross-veins.(O and P) Ectopic expression of Ecto (O), but not of the Ecto RING mutant (Ecto-CAmut [P]), in the wing primordium causes loss of the cross-veins.(Q) Expression of Drosophila Fat facets (Faf) induces ectopic wing veins (red box).(R) Expression of Fat facets antagonizes Ecto, rescuing the formation of the cross-veins.(S and T) Ecto is epistatic to FAM. Panels in (S) show immunoblots of HaCaT cells transfected with the indicated combinations of siRNA. Ecto biological function is required downstream of FAM, as also revealed by TGFβ induced transwell migration assays in MDA-MB231 cells (T). Data are represented as mean and SD. |
