Figure 3
MAPK7 decreases EZH2 through miRNA-101. MiR-101 expression levels were determined by qPCR in HUVEC exposed to FSS (20 dyne/cm2) with or without the MAPK7 inhibitor BIX02189 (10 µM) and normalized to the level of static controls (a). Luciferase reporter binding assays were performed for the 3’UTR of EZH2 in COS7 cells with ectopic expression of miR-101 or scrambled control sequences (scr). Luciferase activity was normalized to non-transfected cells (b). EZH2 and MAPK7 expression levels were determined by qPCR in HUVEC with ectopic expression of miR-101 or SCR and normalized to control (c,e). EZH2 and MAPK7 protein levels were determined by western blot in HUVEC with ectopic expression of miR-101 or scrambled control sequences (d,f). MiR-101 expression levels were determined by qPCR and normalized to IMT < 1 (g). MiR-101 decreases with increasing IMT (h), and associates with MAPK7 (i), and tends to associate with EZH2 (j) expression levels. In coronary artery disease, MAPK7 expression negatively correlates to EZH2 expression (k). Data is expressed as mean ± S.D. of all individual observations. In vitro experimental data was derived from 4 independent experiments, whereas human in vivo data was derived from n = 4–8 samples per group. Comparisons between 2 groups were performed by Student t-tests and data from multiple groups were analyzed by ANOVA followed by Bonferroni post hoc tests. Correlations were performed using Pearson correlation. *p < 0.05, **p < 0.01, ***p < 0.001.
