microRNA 126 Inhibits the Transition of Endothelial Progenitor Cells to Mesenchymal Cells via the PIK3R2-PI3K/Akt Signalling Pathway
Figure 3
PIK3R2 was a direct target of miR-126 in EPCs.
(A) Diagram of PIK3R2-3′UTR-containing luciferase reporter gene construct and the 22-bp target site of miR-126 in PIK3R2-3′UTR. The mutated nucleotides in PIK3R2-3′UTR fragments are underlined. (B) Luciferase reporter assays. The wild-type or mutant reporter plasmids were cotransfected into EPCs, which were infected by control-lentivirus or miR-126-lentivirus. The relative luciferase values shown were normalized to transfections with the wild-type reporter plasmids. Values are the average ± S.D. of 3 replicates. (C)The scheme of the luciferase assay to evaluate the direct inhibition of mir-126 on PIK3R2 protein expression. (D) The expression levels of PIK3R2 mRNA in miR-126 EPCs, miR-126 control, and the negative control—all under TGFβ1 treatment—were examined by qRT-PCR. β-actin was used as an internal control. (E) The protein expression of PIK3R2 was examined by western blotting. β-actin was used as an internal control. ** represents P < 0.01 with an LSD t-test.