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Figure 3. Xbra and Smad-1 synergistically regulate ventx1.1 transcription. Different ventx1.1 promoter constructs were co-injected either with or without Smad-1 and with Xbra, either in combination or separately, at the 1-cell stage and grown until stage 11 to measure the relative promoter activity. (a,b) ventx1.1 (−103 and −180) injected with Xbra and Smad-1, in combination or separately. (c) XbRE-mutated ventx1.1 (−103) mXbRE and ventx1.1 (−103) injected with Xbra and Smad-1, in combination or separately, in separate groups. (d) XbRE and BRE were mutated by site-directed mutagenesis in different ventx1.1 (−180) promoter constructs. (e) XbRE-mutated ventx1.1 (−180) mXbRE injected either with or without Xbra and Smad-1, in combination or separately. (f) BRE-mutated ventx1.1 (−180) mBRE injected either with or without Xbra and Smad-1, in combination or separately. (g) The doubly mutated ventx1.1 (−180) m(BRE + XbRE) injected either with or without Xbra and Smad-1, in combination or separately. (h,i) Flag-Smad-1, Flag-Smad-1 (3SD), and Flag-Smad-1 (3SA) constructs were co-injected with Myc-Xbra and immunoprecipitation was performed with an anti-Flag antibody. (j) ventx1.1 (−180) injected either with or without Myc-Xbra, Flag-Smad-1, and Flag-Smad-1 (3SA) constructs in different groups, in combination or separately. All relative promoter activity data are shown as mean ± SE. Image published in: Kumar S et al. (2018) © The Author(s) 2018. This image is reproduced with permission of the journal 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

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