Figure 4. MiR-221-5p regulates IL-6R expression.
(A) Human IL-6R mRNA contains two predicted miR-221-5p binding sites. (B) A diagram of IL-6R 3′ UTR reporters. (C) WT or mutant IL-6R 3′UTR luciferase constructs were co-transfected with miR-221-5p mimic or control mimic in NCM460-NK-1R cells. Luciferase activity was determined 24 hours after transfection and normalized luciferase activity is shown. *p<0.05, data show mean ± standard deviation of triplicate. (D). MiR-221-5p down regulation increased IL-6R mRNA expression in response to SP stimulation. *p<0.05, data show mean ± standard deviation of triplicate (E). Up-regulation of miR-221-5p decrease IL-6R mRNA expression in response to SP stimulation. *p<0.05, data show mean ± standard deviation of triplicate. (F) IL-6R and β-actin protein levels are assessed by Western blot in miR-221-5p mimic or control-mimic treated NCM460-NK-1R cells.