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. 2010 Jun 28;10:130. doi: 10.1186/1471-2229-10-130

Figure 3.

Figure 3

Binding of EgMYB2 to the regulatory regions of the EgCAD2 and EgCCR promoters containing mutated MYB element(s). (A) EMSAs were performed with recombinant EgMYB2 protein (0.125 μg) and radiolabelled EgCAD2 [-203/-129] or EgCCR [-119/-70] fragments containing individual or multiple mutated MYB elements, as indicated above the lanes by crossed boxes. Lanes 2, 3, 8 and 9 each contain a 100-fold molar excess of unlabelled non-specific (ns, unrelated DNA fragment) or specific (s, EgCAD2 or EgCCR regulatory fragments, respectively) competitor fragment. (B) Transactivation assays were performed by co-transfecting reporter constructs (GUS fused to the -203 EgCAD2 or -119 EgCCR promoters containing the indicated mutated MYB elements) and effector constructs (EgMYB2 cDNA (EgMYB2, hatched bars) or no cDNA (control, open bars) driven by the CaMV 35S promoter) in Nicotiana benthamiana leaves. GUS activity is expressed relative to that driven by the wild-type promoter controls. Histograms represent mean values and standard deviations of two independent experiments, each with three replicates. Activations of EgCAD2 and EgCCR promoters that are statistically significant relative to controls are highlighted by an asterisk (Student's P = 0.01 and 0.003, respectively).