Figure 2
RAN and RAPH1 are direct miR-203 targets. (a) Insertion of miR-203 target 3′-UTRs in a luciferase reporter gene construct leads to a decrease in relative luciferase activity only in the presence of miR-203 target sites (black bars). Deletions of miR-203 target sequences in the RAN and RAPH1 3′-UTRs abolish the miRNA-mediated effect on luciferase activity (gray bars). Histograms represent the average±S.D. from three independent assays. *P-value <0.01 by Student’s t-test. Predicted miR-203 target sites on RAN and RAPH 3′-UTRs. (b) Western blotting of miR-203 targets 48 h after pre-miR-203 or scrambled control sequence transfections. β-Actin expression was used as control. (c) MiR-203 target expression was normalized to actin by densitometry. Results are reported as arbitrary units (AU). (d) Western blotting of miR-203 targets in total protein extracts from dorsal (lanes 1–6) and ventral (lanes 7–12) skin biopsies of new born mice subcutaneously injected on the back side with antagomiR-203 (lanes 1–3 and 7–9) or negative control(lanes 4–6 and 10–12) sequences
