Figure 2

Binding and competition between 3′-UTR of the gene SLC4A4 and miR-224. (A) Lane 1 corresponds to the probe alone (SLC4A4 3′-UTR*) and the other lanes (2–7) correspond to the incubation of the probe with increasing concentrations of miR-224 (1nM-300nM). Lane 8 corresponds to the mismatch probe SLC4A4 3′UTR-MM* alone, lanes 9 and 10 correspond to its incubation with miR-224 (100 and 300nM). The intensity of the bands was quantified with ImageQuant software. Integrated in the panel below, it is represented the increase in the binding between SLC4A4 3′-UTR*:miR-224 (white bars) and the disappearance of the band corresponding to SLC4A4 alone (grey bars). Values correspond to the percentage of intensity of each band with respect to the total radioactivity in each lane. Probes and shifted bands are indicated by arrows. (B) The binding between SLC4A4 3′-UTR* and miR-224 was competed with unlabeled SLC4A4 3′-UTR and SLC4A4 3′-UTR-MM. SLC4A4 3′-UTR* was incubated with miR-224 either in the absence (lane 2) or the presence (lanes 3 and 4) of SLC4A4 3′-UTR and SLC4A4 3′-UTR-MM, respectively (3× the concentration of miR-224). The intensity of the bands was quantified with ImageQuant software. In the panel below, it is represented the corresponding decrease in the binding between SLC4A4 3′-UTR*:miR-224 (gray bars) and the increase of the band corresponding to SLC4A4 3′-UTR* alone (white bars). Values correspond to the percentage of intensity of each band relative to the total radioactivity in each lane.