Fig. 2 Binding between purified recombinant Foxd3 and 32P-radiolabeled oligonucleotide probes was studied with the electrophoretic mobility shift assay. The arrow indicates the shifted band formed by double-stranded oligonucleotide and recombinant Foxd3. Three probes were used: cassette -82/-62, mutated -82/-62 (m4m5, in which the -70/-62 sequence was mutated), and a nonspecific DNA sequence (Non-30fr). Radiolabeled cassette -82/-62, without added nuclear extracts, was the negative control (lane 1). The shifted bands were abolished completely when excess amount of unlabeled -82/-62 oligonucleotide was added (lanes 4 and 5). Recombinant Foxd3 did not bind to m4m5 (lane 8) or Non30fr (lane 9). In addition, m4m5 and Non30fr competitors did not compete for binding with Foxd3 and cassette -82/-62 (lanes 6 and 7).
Reprinted from Developmental Biology, 290(2), Lee, H.C., Huang, H.Y., Lin, C.Y., Chen, Y.H., and Tsai, H.J., Foxd3 mediates zebrafish myf5 expression during early somitogenesis, 359-372, Copyright (2006) with permission from Elsevier. Full text @ Dev. Biol.