Xenbase Image ID: 81442

Fig. 7. Regulation of Sox2, Ngnr1 and Ebf2 by Dmrt5. (A) Animal caps were isolated from embryos injected with Noggin (100 pg), Xenopus MT-Dmrt5 (50 pg) as indicated, cultured until sibling embryos reached stages 14, 22 or 28 and subjected to RT-qPCR analysis. Note that injection of Xenopus Dmrt5 upregulates Ngnr1 and Ebf2 and has no effect on Sox2 in neuralized caps at stages 14 and 22, but not 28. A control western blot shows the decreasing levels of overexpressed MT-Dmrt5 in the caps at the different times analysed. (B) Animal caps from embryos injected with MT-Dmrt5 (50 pg) and Ath5 (250 pg) as indicated and analysed at stage 14 by RT-qPCR for Ngnr1, Ebf2 and Sox2. Note that Dmrt5 slightly induces Ngnr1 and Ebf2 in nae caps and that it promotes their expression in MT-Ath5 overexpressing caps while Sox2 is unaffected. (C) and (D) Animal caps from embryos injected with Noggin mRNA (100 pg) alone or coinjected with mRNA encoding the indicated myc-tagged constructs (50 pg each) harvested at stage 14 and analysed by RT-qPCR for Ngnr1, Ebf2 and Sox2. Note that both the DM and the conserved DMA and DMB domains are required for Dmrt5's ability to induce Ngnr1 and Ebf2 and that mDmrt3, NvDmrtb but not mDmrt2 induces Ngnr1 and Ebf2 expression. In all cases, expression levels were normalized to GAPDH. In (A), (C) and (D), expression levels were compared to the level observed in Noggin injected caps, which was set to 1. Control western blots showing the overproduced proteins are shown. Image published in: Parlier D et al. (2013) Copyright © 2013. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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