Fig. S3. Pkdcc expression is regulated by Wnt and its overexpression induces anterior markers. (A) Schematic representation of the experiment. DMZ (dorsal marginal zone) or VMZ (ventral marginal zone) were explanted from stage 10.5 embryos previously injected at the 4-cell stage with β-catenin morpholino or Xenopus Wnt8 mRNA, respectively. Total RNA was then extracted from the explants, converted into cDNA and analyzed by RT-PCR. (B) Depletion of β-catenin suppressed Pkdcc1 and 2, as well as Chordin (Chd), from DMZ explants. Conversely, Wnt8 overexpression induced ectopic activation of Pkdcc1/2 and Chd transcription in VMZ, while they were not normally transcribed in control VMZ. Stage-matched whole embryos are used as positive control for gene expression, while –RT represents a negative control. H4 is used as a loading control. (C) Quantitative RT-PCR analysis performed on whole embryos injected with β-catenin morpholino or Xenopus Wnt8 mRNA showing induction of Pkdcc. Gene expression levels were normalized to ornithine decarboxylase (ODC) mRNA levels. Error bars are from 3 independent experiments. (D) Xenopus embryos were injected with Pkdcc1 mRNA at 4-cell stage and collected at stage 16 and analyzed by quantitative RT-PCR. mRNA levels of anterior neural markers such as Rx2a, FoxG1, Otx2 and the cement gland marker Xag were increased by overexpression of Pkdcc1. Expression levels were normalized to ornithine decarboxylase (ODC) mRNA levels. Error bars represent 3 independent experiments.
Image published in: Ding Y et al. (2017)
Copyright © 2017. Image reproduced with permission of the Publisher, Elsevier B. V.
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