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. 2017 May 7;8(34):56417–56427. doi: 10.18632/oncotarget.17662

Figure 2.

Figure 2

Dual-luciferase reporter assays for SOS1 (A) and SOS2 (B) as targets for miR-148a. A pSG5 plasmid expressing miR-148a (or the empty pSG5 vector as a control) was co-transfected into HEK-293T cells either with the empty pMIR reporter vector, with pMIR reporter vector containing SOS1 (A) or SOS2 (B) wildtype (WT) 3′UTR, with SOS1 or SOS2 reporter vectors containing a single mutated binding site each (M1, M2) or with pMIR reporter vector for SOS1 (A) or SOS2 (B) containing both mutated binding sites (M1+2) each for miR-148a-3p. Co-transfection of miR-148a with wildtype 3′UTR reporter vectors resulted in relative light units significantly reduced to 77% for both SOS1 (A) or SOS2 (B) reporters as compared to control samples (P < 0.001 each). The downregulation of luciferase activity was abrogated as soon as at least one of the two miR-148a-3p binding sites was mutated.