Figure 2.

miR-193a-3p targets KRas. A. Cells were transfected with either non-targeting miRNA (N), miR-193a-3p (M) or treated with transfection reagent alone (C) for 48 hours. Expression of Ras was determined by western blot. B. Targetscan identifies 2 independent potential binding sites for miR-193a-3p in the 3′UTR of KRas, with no binding sites identified in either HRas or NRas. C. miR-193a shows significant negative correlation with mRNA levels of KRAS (r = −0.11, p = 0.049), but not with either HRAS (r = −0.03, p = 0.57) or NRAS (r = 0.03, p = 0.62). D. Constructs were generated in which the KRas 3′UTR was mutated (×) to prevent miR-193a-3p binding at either the first (S1), the second (S2) or both (B) sites. E. Expression of miR-193a-3p reduces luciferase activity from either the WT, S1 or S2 constructs in A549 cells, but is unable to block activity when both binding sites are mutated (B).