Binding affinity of ERFs to the GCC-box is impacted by the nucleotide composition of the flanking regions. (A) To assess the role of the nucleotide composition of the close environment of the GCC box, nucleotides flanking the chitinase GCC box were mutated. Different mutated GCC boxes were used as probe in gel shift assay to test the binding affinity of ERFs. (B) Binding affinity of ERFs to the mutated probes. Relative affinity is calculated with non mutated Sl-Chitinase (Solyc10g055810.1) as reference. The data are mean of 3 independent replicates. Analysis of variance with the R package reveals that the flanking region of the GCC box is significantly involved in the affinity of the binding (p<0.05).