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Research Paper Volume 11, Issue 3 pp 908—920

Figure 2. MiR-449 targets 3’-UTR of PLOD1 mRNA to inhibit its protein translation in ASCs. (A) Rat ASCs cells were transfected with plasmids carrying miR-449 or as-miR-449 or scr as a control. RT-qPCR for miR-449 was done. (B) The intact 3'-UTR of wildtype PLOD1 mRNA (wt PLOD1 3'-UTR) and the 3'-UTR of PLOD1 mRNA with a mutant at miR-449-binding site (mut PLOD1 3'-UTR) were respectively cloned into luciferase reporter plasmids. Rat ASCs cells were then co-transfected with one plasmid from miR-449/as-miR-449/null plasmids and one plasmid from either wt PLOD1 3'-UTR or mut PLOD1 3'-UTR, and subsequently subjected to a dual luciferase reporter assay. (C) RT-qPCR for PLOD1 mRNA in miR-449-modified rat ASCs. (D) Western blotting for PLOD1 protein in miR-449-modified rat ASCs. *p<0.05. NS: non-significant. N=5.