Xenbase Image ID: 147671

Supplemental Figure S2 (A) Ecto-depleted animal cap cells express mesoderm specific markers to levels comparable to those triggered by Smad4 mRNA injection (2 ng). Activin mRNA injection (20 pg) triggers stronger inductions. Quantitations of VegT inductions: Ecto- MO, 1X; Smad4, 2X; Activin, 32X. (B) Targeted depletion of Ectodermin from the prospective mesoderm leads to ectopic expression of endoderm markers. Eight-cell stage embryos were injected either in the marginal zone (MZ) or in the vegetal pole (Vg) with 60 ng of control-MO (-) or Ecto- MO (+) together with GFP mRNA as a tracer. At early gastrula, GFP-positive cells were dissected under the GFP-scope using Gastromaster wires (left panel shows representative explants before hasrvesting). RT-PCR analysis for Mixer and Mix.2 shows a specific effect of Ecto-knockdown in mesoderm cells (MZ), but not in vegetal explants (Vg) where Ectodermin is not expressed. RT-PCR analysis for Xbra and Sox17α proves absence of cross-contaminations in the explants. The effect of Ecto-MO in MZ explants was validated by immunoblot and found to be quantitative (data not shown). Image published in: Dupont S et al. (2005) Copyright © 2005. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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