Figure 2

Aberrant expression of miR-133a in endothelial cells alters the expression of key regulators of DLL4-Notch signalling. (A) Volcano plot representation showing differential gene expression analysis of a RT2 Profiler PCR Array Human Notch Signalling Pathway Plus Kit (Ref PAHS-059Y, Qiagen) using RNA isolated from HUVEC transfected with miR-133a-3p or negative control (miR-NC) and stimulated for 1 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–I) 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 the indicated times. 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 = 9. Data were analysed for statistical differences by two-way (B–G) or one-way (H–I) ANOVA with post hoc Tukey’s comparison test. ns = non-significant; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001.