Binding site specific mutagenesis prevents the modulation of <i>ACVR1/Alk-2</i> expression operated by the selected miRs.

Abstract

<p> Mutagenesis of miRs binding sites was introduced in the two seed sequences predicted for mir148b (A) and mir365 (B), indicated as Seed1 and Seed 2, and in the mir26a seed sequence (C). Site specific mutagenesis was also performed to abolish the first AUUUA element contained in the mir26a pairing region (ATTTA mut), alone or in combination with the mutation of the mir26a seed sequence (double mutant mir26 mut/ATTTA mut). Each pair of bars represents Luciferase activity in the presence of negative control miR (-), considered as 100%, or the specific miR (+). <i>Student’s t-test</i> was applied to evaluate differences between the effect of specific mir compared to that of the negative control for each construct, <i>p</i><0.05*, <i>p</i><0.01**, <i>p</i><0.001***.</p

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