Xenbase Image ID: 154068

Fig. 4. Tbx2 binds to the promoter of Flrt3 gene. (A) Alignment of wild-type (wt) putative T-box binding sites (in red) in the promoter regions of Xenopus and human Flrt3 and the sequence of the mutant (mut) binding sites. (B) Chromatin immunoprecipitation (ChIP) analysis using HEK 293 T cells transfected with human Tbx2 along with DNA constructs harboring wild-type or mutant T-box binding sites of human Flrt3. P1, P2 and CP denote primers used for PCR reactions (Materials and Methods). Input, promoter DNA in cross-linked DNA-protein mix before immunoprecipitation. Ab (-), immunoprecipitation without antibody. Co. IgG, control rabbit IgG. (C) Suppression of the activity of Flrt3 promoter-driven reporter by Tbx2 or BMP4 requires T-box binding sites. Embryos were injected dorsally with pGL3-wt Flrt3pro-luc (40 pg), pGL3-mut Flrt3pro-luc (40 pg), LacZ (200 pg), Tbx2 (200 pg) and BMP4 (200 pg) as indicated and incubated until stage 11, when firefly and Renilla luciferase activities were determined. Each experiment was carried out in triplicate and error bars indicate the standard error (SE) from three independent experiments. *p-value <0.05 and **p-value <0.01. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.) Image published in: Cho GS et al. (2017) Copyright © 2017. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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