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. 2007 Jun 25;104(27):11310–11315. doi: 10.1073/pnas.0611299104

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© 2007 by The National Academy of Sciences of the USA

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Fig. 2. — The MEF3/Six binding site present in the 145-bp sequence is conserved between species and binds Six1 and Six4 proteins. (A) DNA sequence alignments, from different species, of the 145-bp Myf5 element (26), showing 100% sequence conservation of the putative MEF3 sites. MEF3 and Pax3 sequences are underlined. The MEF3 consensus is shown below the MEF3 box site. (B) Electromobility shift assays with a 30-bp oligonucleotide containing the Myf5 MEF3 site (lanes 1–11) or a mutant MEF3 site (lanes 12 and 13), incubated with recombinant Six1 protein (lanes 2–11 and 13) or with crude reticulocyte lysate (lanes 1 and 12). Six1 antibodies (lane 11) or a 66- or 200-fold excess of MEF3-Myf5 (lanes 3 and 4), MEF3-Myogenin (lanes 5 and 6), mutant MEF3-Myf5 (lanes 7 and 8), or NF1-Myogenin (lanes 9 and 10) unlabeled competitors was added in the mix. Note that Six1 is unable to recognize the mutant MEF3 oligonucleotide.