Figure 2.

miR-133a putative target analysis. (A) The putative targets of miR-133a were analysed by means of the TargetScan-7.1 software and here are represented the consensus sequence and the conservation across different species [45]. A conserved putative binding site was retrieved in the 3′ UTR of the TPM4 transcript. (B) A 300 bp fragment, containing the putative responsive element, was cloned into the pGL3 vector, replacing part of the 3′ UTR of the luciferase gene, this will permit to control the luciferase gene if the cloned sequence contains a genuine binding site for miR-133a. (C) Assay showing the inhibition of luciferase when the vector is co-transfected with miR-133a demonstrates its interaction with the cloned responsive element (n = 6, * p < 0.01, t-test two tails, coupled). (D) miR-133a amplification from RNA extracted from plasma and from isolated exosomes. (n = 3).