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. 2009 Oct 9;136(21):3679–3689. doi: 10.1242/dev.038141

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Fig. 3. — In vitro binding of the sequences necessary for notochord activity of the Ci-tune CRM by the Ci-Bra and Ci-FoxA-a proteins. (A) Electrophoretic mobility shift assays (EMSA) with a radiolabeled probe containing the predicted Ci-FoxA-a binding site and a GST-Ci-FoxA-a fusion protein. Lanes contained: (1,2) radiolabeled probe containing the predicted Ci-FoxA-a binding site, (1) free, and (2) incubated with the GST-Ci-FoxA-a fusion protein; (3,4) radiolabeled probe containing the mutated Ci-FoxA-a binding site, (3) free, and (4) incubated with GST-Ci-FoxA-a; (5-12) radiolabeled probe containing the predicted Ci-FoxA-a binding site and unlabeled competitors, at 1 μM (lanes 5,7,9,11) and 5 μM (lanes 6,8,10,12) concentrations, incubated with GST-Ci-FoxA-a. (B) EMSA with radiolabeled probe and a GST-Ci-Bra fusion protein. Lanes contained: (13,14) radiolabeled probe containing the predicted Ci-Bra binding site T-box site 1, (13) free, and (14) incubated with GST-Ci-Bra fusion protein; (15-18) radiolabeled T-box site 1 probe and unlabeled competitors, at 1 μM (lanes 15,17) and 5 μM (lanes 16,18) concentrations, incubated with GST-Ci-Bra.