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Figure 3


Fig. 3. Rspo3 acts via Wnt/β-catenin signaling to induce Vegf. (A-C) RT-PCR analysis or quantitative PCR analysis (qPCR) of indicated marker genes in ventral marginal zones (VMZ). Four-cell stage Xenopus embryos were injected with 2.5 ng of Control Mo or Rspo3 Mo, 10 ng of VEGF Mo, 50 pg Xenopus Rspo2 mRNA, 250 pg dominant-negative Wnt8 (dnWnt8), 50 pg pCSKA-Wnt8 DNA, 150 pg Xenopus Dkk1 DNA, 50 pg Xenopus Rspo3 mRNA or 250 pg Xenopus VEGF-A170 mRNA, as indicated, into each ventral blastomere. VMZs were explanted at stage 10.5, cultured until stage 28 with or without treatment of VEGF receptor inhibitors (KRN or MAZ), and processed for RT-PCR (A,B) or qPCR (C). -RT, minus reverse transcription control. (D) Four-cell stage embryos were injected with 2.5 ng of Control Mo or Rspo3 Mo, as indicated, into both ventral blastomeres. Embryos were cultured until the indicated stage and prospective vbi regions were dissected for qPCR analysis. Gene expression in uninjected vbi explants was set to 100% (control).





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