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. 2017 Jul 15;144(14):2629–2639. doi: 10.1242/dev.146241

Fig. 6.

Fig. 6.

The notch1b-15 enhancer contains essential SoxF binding motifs. (A) Multispecies alignment of the orthologous regions of the notch1b-15 enhancer from zebrafish (zfish), fugu, stickleback (stickle) and medaka using ClustalW. Coloured sequences are confirmed by EMSA; grey sequences are motifs identified in silico that did not bind robustly in EMSA. (B) Radiolabelled oligonucleotide probes encompassing notch1b-15 zfSOX-a (lanes 1-8) and zfSOX-b (lanes 9-16) were bound by recombinant SOX7 (lanes 2-4 and 10-12) and SOX18 (lanes 6-8 and 14-16). Both proteins, which efficiently bound labelled probes (lanes 2, 6, 10 and 14), were competed by excess unlabelled self-probe (WT, lanes 3, 7, 11 and 15) but not by mutant self-probe (MT, lanes 4, 8, 12 and 16). (C) Radiolabelled oligonucleotide probes encompassing notch1b-15 zfETS-a (lanes 17-20) and zfETS-b (lanes 21-24) (see A) were bound by recombinant ETV2 proteins. ETV2, which efficiently bound to labelled probes (lanes 18 and 22), was competed by excess unlabelled self-probe (WT, lanes 19 and 23) but not by mutant self-probe (MT, lanes 20 and 24).