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. 2015 Oct 12;16(10):24032–24047. doi: 10.3390/ijms161024032

Figure 3.

Figure 3

MiR-30a Targets the 3ʹUTR of NFATc3 and Inhibit the Expression of NFATc3. (A) Schematic presentation of two binding sites of miR-30a in the 3ʹUTR of NFATc3 and NFATc3 3ʹUTR reporter constructs, including wild type and mutant vectors; (B) Dual-luciferase reporter assays demonstrate that the wild-type and mutation-BS2 reporter of NFATc3 3ʹUTR suppressed activity in HEK293T cells and MPC5 cells transiently transfected with miR-30a, but not in mutation-BS1 and mutation-BS 1 and 2; (C) RT-qPCR and (D) Western blot analysis of NFATc3 expression levels in cultured MPC5 cells transfected with miR-30a (30 nM) or miR-NC. Quantitative analysis of NFATc3 protein level after normalization with β-tubulin; (E) RT-qPCR and (F) Western blot analysis of NFATc3 expression levels in cultured MPC5 cells transfected with inhibitor-30a (50 nM) or inhibitor-NC. Quantitative analysis of NFATc3 protein level after normalization with β-actin. The relative amount of NFATc3 mRNA was normalized to 18 s. All data are presented as means ± SD from three independent experiments. * p < 0.05, ** p < 0.01, indicate highly statistically significant differences, compared with miR-NC or inhibitor-NC, respectively.