Figure 3.

TCF7L2 is an miR-26b-5p target gene. (A) Aligned miR-26b-5p sequences from different species. (B) Sequence complementarity between miR-26b-5p and the TCF7L2 3′-UTR. (C) A luciferase reporter assay was used to confirm interactions between miR-26b-5p and TCF7L2, revealing that miR-26b-5p mimic transfection significantly reduced the activity of the WT but not the MUT reporter construct relative to cells transfected with a control. (D) A second luciferase reporter assay revealed that miR-26b-5p inhibitor transfection was associated with a significant increase in the activity of the WT but not the MUT TCF7L2 3′-UTR reporter construct. (E) The expression of TCF7L2 was assessed by quantitative polymerase chain reaction in hADMSCs following miR-26b-5p mimic or inhibitor transfection, resulting in significant decreases and increases in this expression, respectively, relative to cells transfected with control constructs. (F) TCF-4 protein levels were assessed by western blotting in hADMSCs transfected with miR-26b-5p mimics and inhibitors, revealing significant decreases and increases, respectively, in these protein levels relative to cells transfected with control constructs. GAPDH: glyceraldehyde 3-phosphate dehydrogenase; hADMSC: human adipose-derived mesenchymal stem cell; TCF-4: T cell factor 4; UTR: untranslated region; WT: wild type.