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. 2007 Feb 28;2(2):e253. doi: 10.1371/journal.pone.0000253

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Fic et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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In vitro DNA binding activities of Ey and Ey(2a) paired domains. (A) SDS-PAGE of GST-PD and GST-PD(2a) fusion proteins that were expressed and purified from bacteria. (B) EMSA performed with GST-PD (lanes 1–4) or GST-PD(2a) (lanes 5–8) in the presence of four established consensus sequences P6CON (lanes 1 and 5), 5aCON (lanes 2 and 6), CD19-2 (lanes 3 and 7) and sowt3 (lanes 4 and 8) for Ey or its human homolog PAX6 or PAX6(5a) isoform. See text for details. (C) EMSA of P6CON (left panel) and sowt3 (right panel) probes in the presence of increasing amounts of GST-PD (1 ng, lane 1; 5 ng, lane 2; 50 ng, lane 3; 200 ng, lane 4) and GST-PD(2a) (1 ng, lane 5; 5 ng, lanes 6, 50 ng, lane 7; 200 ng, lane 8) proteins. (D) EMSA of P6CON (left panel) and sowt3 (right panel) probes with 50 ng of GST-PD (lanes 1–4) or GST-PD(2a) (lanes 5–8) in the presence of (1 fold, lanes 2 and 6; 2 fold, lanes 3 and 7; 10 fold, lanes 4 and 8) of unlabelled corresponding probes or without competitor (lanes 1 and 5).