XB-IMG-76661
Xenbase Image ID: 76661
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Fig. S1. Meis3 expression is repressed by neural Meis3-induced Wnt3a. (A) Scheme of the Meis3/NE recombinant explant experimental design. Meis3 mRNA (0.75 ng) overexpressing pigmented AC explants were recombined with neuralized AC ectoderm (NE). ACs were neurally induced by injecting embryos with BMP DNR mRNA (175 pg); Wnt3a-deficient NE, was co-injected with the Wnt3a MO (45 ng). Explants were cultured and Meis3 expression was examined at late gastrula or mid-neurula stages. (B) Results of the experiment in A. Meis3 expression is expanded in Wnt3a-deficient NEs. Expression of Wnt3a in the NE is dependent on Meis3 from the juxtaposed AC explant (not shown). Thus, neural, Meis3-induced Wnt3a represses Meis3 expression. The dashed line indicates the border between the Meis3 (left) and the NE (right) explants. The pigmented Meis3 AC is strongly over-stained by the Meis3 probe. (C) The experiment was performed as in A, but injections were Meis3 (0.5 ng), BMP DNR (160 pg) and Dkk1 (35 pg) instead of the Wnt3aMO. After culturing recombinants to mid-late neurula stages, the elongated albino NE explant-side was dissected and lysed for RT-PCR to hindbrain markers. Isolated NE-AC, not recombined with Meis3-AC serves as a control. While hindbrain marker expression was typically increased, the HoxB9 spinal cord marker is decreased as a result of Dkk1 activity. (D) RT-PCR to hindbrain markers in mid-neurula stage AC explants from embryos injected with Meis3 mRNA (250 pg), Meis3/Dkk1 (35 ng), or Meis3/Wnt3aMO (40 ng). (E) RT-PCR to Krox20 in mid-neurula stage neuralized AC explants from embryos injected with Noggin mRNA (10 pg) and Meis3 (400 pg) mRNA, or noggin/Meis3/Dkk1 (35 ng), or noggin/Meis3/Wnt3aMO (40 ng). 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. Larger Image Printer Friendly View |