Figure 3

Interaction of miR-708 with the 3’UTR of CD38. A: Predicted binding sites for miR-708 and miR-140-3p in 3’UTR of CD38. Shown are the mutated bases in the miR-708 target site of the 3’UTR of CD38. B: Relative luciferase activity in NIH-3 T3 cells co-transfected with reporter plasmid and different concentrations of miR-708 mimic (mimic) or scrambled sequence mimic (scr) at the highest concentration (n = 3, experiments in triplicate). Note significant reduction in luciferase activity following transfection with miR-708 mimic at all concentrations. C: Relative luciferase activity in cells co-transfected with reporter plasmid containing either wild-type (wild) or mutant (mut) CD38 3’UTR followed by transfection with 50 nM of miR-708 mimic (mimic) or scrambled sequence mimic (scr) (n = 3, experiments in triplicate). Note no inhibition of luciferase activity in cells co-transfected with mutant CD38 3’UTR. D: CD38 mRNA stability measured in growth-arrested HASM cells. Cells were transfected with 50 nM miR-708 and exposed to rh-TNF-α in the presence of actinomycin D. At time points indicated, CD38 transcript levels were quantified by q-PCR (n = 2–5 donors). Graph shows plots of non-linear regression of mRNA decay over time for NA-HASM and AS-HASM cells, with the dotted lines indicating the best fit. Values are means ± SEM for results shown.