XB-IMG-41550
Xenbase Image ID: 41550
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Fig. 6. Meis3 is a direct target of β-catenin. (A) CHX AC assay (Materials and methods). sqRT-PCR of pools of 18 mid-late gastrula ACs were dissected from embryos injected with 20 pg THVGR mRNA at the one-cell stage. ACs were treated with CHX, DEX or CHX+DEX. Ef1α expression serves as a positive control for CHX activity and ODC as a loading control (n=2 experiments). (B) The XMeis3 genomic locus. Green ovals indicate β-catenin/TCF-binding sites, the blue line indicates the ChIP enriched fragment detected in C; the pink line indicates the 2.7 kb used for the constructs in D and E. (C) ChIP for β-catenin in late-gastrula embryos. A region 1.8 kb upstream of the Meis3 transcriptional start site is specifically enriched. Data are plotted as mean fold enrichment by β-catenin IP (black bars) over control serum IP (gray bars) for Meis3 (–1813) or a negative control locus XMLC2 (–118). Error bars show s.e.m. (n=3). (D) Reporter constructs used in E. The 2.7-luc construct contains wild-type β-catenin/TCF-binding sites (green ovals), whereas these are mutated in the 2.7δTCF-luc construct (red ovals). (E) Meis3 promoter activity is dependent on two β-catenin/TCF-binding sites. In situ hybridization of early- and mid-neurula 2.7-luc (b,e) and 2.7δTCF-luc (c, f) transgenic embryos, and of wild-type embryos (a,d). Image published in: Elkouby YM et al. (2010) Copyright © 2010. 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 |