Figure 6. WNT3 interacts directly with miR-637. (A) qRT-PCR analysis of relative WNT3 and β-catenin expression in PBMNCs of hypertensive subjects and normotensive controls. (B) Western blot analysis of WNT3 and β-catenin levels in plasma of hypertensive subjects and normotensive controls. (C) Bioinformatics evidence of the interaction between miR-637 and the 3′-UTR of WNT3. Bottom: schematic diagram of the mutations in the WNT3 sequence used to create the mutant luciferase reporter construct. (D) Luciferase activity assay in HEK293T cells co-transfected with miR-637 mimics or mimic NC and luciferase report plasmids containing wild type (wt) or mutant (mut) WNT3 3′ UTR and. (E) RIP assay results showing co-precipitation of miR-637 and WNT3 mRNA complexes by an Ago2 antibody in HASMCs. Validation data obtained by qRT-PCR are also shown. (F) Western blot analysis of WNT3 levels in HASMCs transfected with miR-637 mimics or mimic NC. Expression data are normalized to GAPDH. (G) Spearman's correlation analysis of the relationship between LOC646616, WNT3, and miR-637 levels, and between WNT3 and β-catenin levels in our hypertensive cohort. Data are presented as the mean ± SD. n = 3 biologically independent samples. *P<0.05, **P<0.01.