Differential expression of microRNAs following cardiopulmonary bypass in children with congenital heart diseases

Fig. 1. Unsupervised hierarchical clustering (Euclidian distance, complete linkage) of the three patients (six samples, three before and three after CPB) based on expression of the 50 with highest variance. The heatmap shows miRNAs with high expression in blue, miRNAs with low expression in red. The blue and orange lines indicate the two main clusters of samples

Fig. 2. Relative fold change of the miRNAs in the atrial myocardial tissue of patients with CHD after CBP (n = 11) compared to before CPB (n = 11) as determined by RT-qPCR and after CBP (n = 3) compared to before CPB (n = 3) as determined by microarray (P < 0.05). Paired-two-tailed t-tests and ±standard deviation (STDV) were used to evaluate differences in expression in both assays. Relative expression level of 2−ΔΔCt and quantile normalization were used for RT-qPCR and microarray data, respectively

Fig. 3. Validation of nine differentially expressed miRNAs in the atrial myocardial tissue of patients with CHD after CBP (n = 11) compared to before CPB (n = 11) as determined by RT-qPCR (P < 0.05). Mean ΔCt before CPB and after CPB (Lower ΔCt, higher expression level). RNAU6B as an endogenous control for normalization, paired-two-tailed t-tests and ±standard deviation (STDV) were used to evaluate differences in expression

Fig. 4. Significantly expressed miRNAs in the atrial myocardial tissue of neonates and infants with CHD after CBP compared to before CPB as determined by RT-qPCR (P < 0.05). Mean ΔCt before CPB and after CPB (Lower ΔCt, higher expression level). RNAU6B as an endogenous control for normalization, paired-two-tailed t-tests and ±standard deviation (STDV) were used to evaluate differences in expression

Fig. 5. Target network with deregulated miRNAs indicated in brown and target genes in blue. Only genes targeted by at least two or ore miRNAs are included. Strong interaction was observed between miRNAs and genes are highlighted by “green” edges in the resulting network