Figure 5. MicroRNA expression profiling in ischemic CMECs proliferation(ICP), ischemic CMECs tube formation (ICTF)and ischemic CMECs migration (ICM)compared with that of normal CMECs proliferation(NCP), normal CMECs tube formation (NCTF) and normal CMECs migration (NCM).

A. Differentially expressed miRNAs in ICP(n = 3), ICTF(n = 3) and ICM(n = 3) compared with NCP(n = 3), NCTF(n = 3) and NCM(n = 3). The heat map diagram showed the result of the two-way hierarchical clustering of miRNAs and samples. Each row represented a miRNA and each column representsed a sample. The miRNA clustering tree was shown on the left, and the sample clustering tree appeared at the top. The color scale shown at the top illustrated the relative expression level of a miRNA in the certain slide: red color represented a high relative expression level; green color represented a low relative expression levels. n = 3 in each group. B. Volcano Plots were useful tool for visualizing differential expression between two different conditions. They were constructed using fold-change values and p-values, and thus allowing visualizing the relationship between fold-change (magnitude of change) and statistical significance (which took both magnitude of change and variability into consideration). They also allowed subsets of genes to be isolated, based on those values. The vertical lines corresponded to 1.5-fold up and down, respectively, and the horizontal line represented a p-value of 0.05. So the red point in the plot represented the differentially expressed miRNAs with statistical significance. C. Confirmation of mir-223-3p in the same set of samples as used in microarray assay by means of real-time PCR. ΔCt values were normalized to U6 levels. Relative expression was calculated with respect to normal CMECs. The results were expressed as Log10 (2∧^−ΔΔCt). *P<0.05, **P<0.01.