Computational analyses allow refinement of functional Svb binding sites. (a) Statistical analysis of positive enhancers versus negative regions (intergenic genomic sequences used as background) was performed for de novo discovery of motifs, showing evolutionary conservation across Drosophila species and characteristics of active enhancers. (b) The svbF7 and blue motifs perform best in discriminating between positive enhancers and negative regions as illustrated by the Pareto plot. (c) While disruption of the core CnGTT OvoQ6 motif abolished Emin activity, point mutations that affect the 5' and 3' flanking nucleotides strongly reduced epidermal expression, as shown by anti-lacZ immuno-staining and quantification of fluorescence signals. wt, wild type. Error bars represent the standart deviation. *** P-value < 0.001; ** P-value < 0.01.