Figure 5

Ectopic expression of miR-133a-3p alters the expression of key mediators of ECM remodelling. (A) Volcano plot representation showing differential gene expression analysis of a RT2 Profiler PCR Array Human Extracellular Matrix & Cell Adhesion Molecules Kit (Ref PAHS-013Z, Qiagen) using RNA isolated from HUVEC transfected with miR-133a-3p or negative control (miR-NC) and stimulated for 4 h with VEGF-A₁₆₅ (50 ng/ml). Results of three independent experiments were analysed using GeneGlobe Data Analysis Center (Qiagen). Genes showing a Log2(miR-133a-3p/miR-NC) > 1 and − Log10(P value) P ≤ 0.05 were selected as upregulated (red). Those with Log2(miR-133a-3p/miR-NC) < − 1, and − Log10(P value) P ≤ 0.05 were selected as downregulated (green). (B–E) HUVEC transfected with mimics for negative control (NC, grey bars), miR-133a-3p (3p, blue bars) or miR-133a-5p (5p, red bars) were left unstimulated (0 h) or stimulated with VEGF-A₁₆₅ (50 ng/ml) for 4 h. RNA isolated from these cells were used to determine changes in gene expression using qPCR TaqMan Gene Expression Assays specific for each gene. Data are shown as mean ± SE, n = 8. Data were analysed for statistical differences by two-way ANOVA with post hoc Tukey’s comparison test. ns = non-significant; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001.