Xenbase Image ID: 76510

Fig. 2. Wnt3a represses Meis3 expression via auto-regulation by Meis3 protein. (A)Meis3-5'UTR (a,b) and Meis3-ORF (c,d) expression in mid-neurula embryos injected at the one-cell stage with high levels of Meis3 mRNA (1 ng). (B)Meis3-5'TR expression in mid-neurula embryos injected at the one-cell stage with Wnt3 CMV-driving plasmid (60 pg; pCMV:Wnt3; b) or Meis3-MO (30 ng; c), or both (d). (C)RT-PCR to endogenous Meis3 (Meis3-5'UTR primers) in AC explants from embryos injected with THVGR mRNA (50 pg) or Meis3-MO (30 ng), or both. RNA was isolated from a pool of 18 mid-neurula explants in each group. Results were quantified, normalized to Ef1-alpha loading control and plotted as fold-repression. Bars are s.e.m. from three independent experiments. (D)CHX assay and RT-PCR in embryos (lanes 1-2) and AC explants (lanes 3-7) injected with Meis3-GR mRNA (250 pg). At early gastrula stages, 5uM CHX was added to explant medium 2 hours prior to Dex activation (1uM). Explants were analyzed at late gastrula stages. Unactivated Meis3-GR shows no leaky activity. Ef1-alpha is a positive control for CHX activity, ODC is a loading control. T samples showed no DNA contamination. (E)A schematic summary of the results. Image published in: Elkouby YM et al. (2012) Copyright © 2012. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Gene Synonyms Species Stage(s) Tissue meis3.L meis3-a, meis3-b, mrg2, XMeis3 X. laevis Throughout NF stage 18 hindbrain midbrain neural plate Image source: Published Larger Image Printer Friendly View

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