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Fig. S1. DNA sequence analysis of the myomaker promoter in multiple animals to find putative NFAT binding sites.
The sequence 1500 bp upstream of the (a) squirrel and mouse, and (b) squirrel and human myomaker transcriptional
start sites were interrogated using rVista for the detection of NFATc1-4 binding sequences. Red lines indicate
aligned hits of possible NFATc1-4 binding regions between organisms, the two sequences highlighted were
identified by rVista as possible binding sites. Cross-referencing with NFAT binding sequences reported in the
literature (c, shown in green), one probable binding site remained 1095 bp upstream of the myomaker transcriptional
start site. This sequences along with its aligned flanking region from the multiple alignments was used to design a
25 bp probe for DNA ELISA.
Fig. S2. (a) Amino acid multiple alignment using ClustalW2 for myomaker proteins from human, mouse, and
squirrel showing strong sequence conservation. * indicates identical residues, : indicates conservation between
amino acids of similar properties and . represents dissimilar properties. Red letters represent hydrophobic amino
acids, blue – acidic, magenta – basic, green – hydroxyl, sulfhydryl, or amine. (b) Pair-wise amino acid alignment
percent identities were determined using ClustalW2 for squirrel, mouse, and human NFATc1-4, myoferlin, and
myomaker proteins.