Xenbase Image ID: 84309

Figure 4. Rspo2 Inhibits TGF-β Signaling in Xenopus. (A) 4-cell stage embryos were injected animally with 100 pg Rspo2, 100 pg Xwnt8, or 200 pg β-catenin mRNA in each blastomere. At stage 8, animal caps were dissected, cultured until stage 18 equivalent, and analyzed for expression of the indicated marker genes. −RT, minus reverse transcription control. (B) 2-cell stage embryos were injected with 100 pg Rspo2 or preprolactin (ppl) mRNA in each blastomere. Note ectopic cement glands (arrowhead) and shortened body axis in Rspo2-injected embryo. (C-J) Whole-mount in situ hybridizations of the indicated genes. 8-cell stage embryos were injected with 100 pg of Rspo2 or ppl mRNAs as indicated into one animal blastomere. LacZ mRNA was coinjected as lineage tracer in all panels except (C), (G), and (H). (C-H) Stage 15 neurulae in anterior view. (I and J) Stage 11 gastrulae in vegetal view, dorsal up. (K-N) 4-cell stage embryos were injected animally with indicated RNAs. At stage 8, animal caps were dissected, cultured until stage 10 equivalent, and analyzed for expression of Xvent2 (K) or Xbra (L-N). −RT, minus reverse transcription control. Embryos injected with 100 pg preprolactin (ppl) were used as control. Amounts of mRNAs used: 100 pg Rspo2, 50 pg or 250 pg of BMP-4, 50 pg of activin, 25 or 100 pg of Wnt8, 200 pg of β-catenin, 50 or 100 pg of Xnr1, 2 or 20 pg of FGF8. (O) BMP responsive luciferase reporter assay in 293T cells. Concentrations of plasmids were: BRE4-luc, 10 ng, BMP4, 10 ng, Wnt1, 5 ng, Rspo2, 5 ng. Image published in: Kazanskaya O et al. (2004) Copyright © 2004. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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