Xenbase Image ID: 49548

Figure 5. Chordin Is Required for the Induction of Dorsal Mesoderm by Activin(A–F) Two-cell embryos were injected four times into the animal pole with a total of 8 ng of Chd-MO-1, Chd-MO-2, Chd-MO(1+2), or with Chd-MO(1+2) and 10 pg of chordin mRNA lacking the 5′ UTR of the chordin cDNA. Animal explants were isolated at stage 8, treated with 2 ng/ml Activin, and cultured until stage 25. Three independent experiments were performed with similar results.(G) RT-PCR analysis of samples shown in (A–F). Activin treatment induced the expression of the somite marker genes Myf5 and Actin and the pan-neural marker NCAM. Chd-MO(1+2) inhibited the expression of dorsal markers but upregulated the expression of ventral marker genes (xHox3 and α-Globin).(H) A subset of the animal caps were cultured overnight in CMFM. The supernatant was analyzed in Western blots with an immunopurified antibody specific for the interrepeat region of Chordin (α-I-Chd; Larraı́n et al., 2001), and the cell pellet was analyzed with an antibody specific for phospho-Smad-1.(I) RT-PCR analysis of Activin-treated animal caps at stage 10.5 injected with Chd-MO(1+2). The morpholinos did not affect mesoderm induction (Xbra) or the expression of other BMP antagonists (cerberus, chordin, noggin, and follistatin).(J) RT-PCR analysis of Activin-treated animal caps at stage 12. The microinjection of Chd-MO(1+2) changed noggin and follistatin mRNA levels only slightly, inhibited chordin expression, and upregulated the ventral mesoderm marker xHox3. Image published in: Oelgeschläger M et al. (2003) Copyright © 2003. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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