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Fig. 3. Smad7 blocks Smad1 and Smad2 phosphorylation but induces neural markers only indirectly. (A and B) Inhibition of phospho-Smad1 and phospho-Smad2 by Smad7, tAlk4+Smad6 and CerS+Smad6, revealed by Western blot analysis of animal caps from injected embryos. Fldx-injection and whole embryos are included as controls. (B) Quantification of the level of phosphorylated Smad1 and Smad2 in the above experiment. (C) Effects of decreasing concentration of Smad7 on Sox3, Sox2 and Chordin induction after injection into the A4 blastomere. 2 ng: Sox3 92%, Sox2 81%, Chordin 100%; 1–0.5 ng: Sox3 89%, Sox2 80%, Chordin 100%; 100 pg: Sox3 66%, Sox2 36%, Chordin 91%; 50 pg: Sox3 33%, Sox2 14%, Chordin 88%; 10 pg: Sox3 0%, Sox2 0%, Chordin 72%. (D) The induction of Sox3 is not cell-autonomous to the progeny of the Smad7-injected A4 blastomere. Embryos were injected with either Rldx or Smad7+Fldx in the A4 blastomere at the 32-cell stage, and isotopic transplants performed at stage 8, replacing Rldx-labelled cells with the progeny of Smad7(2 ng)+Fldx-injected cells. Embryos were grown to stage 18 and stained for Sox3 and Fldx. (E) Section through the transplanted area showing expression of Sox3 in uninjected cells of the host (8/8). Image published in: de Almeida I et al. (2008) Copyright © 2008. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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