Fig 1.
Heatmap displaying the 153 most differentially expressed probes (p<0.05 FC>1.2) in the RV Endocardium of 5 controls pigs (T1 to T5) and of 7 rTOF pigs (F1 to F7).
The red and green colors indicate relative transcript abundance (red = overexpressed, green = downregulated). The columns represent the 12 samples while the rows correspond to the 153 probes. Samples were classified using hierarchical clustering, according to similarity in change in relative transcript abundance.
Fig 2.
The significant gene ontology biological processes in the RV Endocardium.
Fig 3.
Functional categorization analysis of the most significant pathways represented in the microarray-generated list of the 54 most differentially expressed genes (Ingenuity software) in the RV Endocardium of rTOF pigs.
The p-values (blue bars) were calculated using the right-tailed Fisher’s exact test. The threshold (yellow line) is set to a p-value = 0.05. The ratios (yellow dots and curve) represent the number of molecules of the experiment related to the number of total molecules involved in the pathways.
Fig 4.
A. Ingenuity Pathway Analysis (IPA) generated 4 molecular networks assembled from DE genes the RV endocardium of rTOF pigs versus Controls (genes in bold) in the RV Endocardium. B. IPA Network #2. based upon differentially expressed genes that were upregulated and down regulated in the RV Endocardium.
Fig 5.
A. Relative expression (RT-qPCR) of genes in samples from the RV Endocardium of controls (grey bars) and rTOF (black bars) hearts. Transcript expression is normalized to the reference genes HPRT1 and GUSB. Two sided T-Test Statistical significance of n = 5 Controls and n = 7 rTOF hearts (*P <0.05, **P <0.01, ***P <0.001) B. Fold change comparison in samples from the RV Endocardium (n = 5 Controls and n = 7 rTOF hearts) based on RT-qPCR results and microarray expression data. Pearson correlation coefficient R = 0.77 (p = 0.001).
Fig 6.
Relative expression (RT-qPCR) of BNP, Serca2a (ATP2A2) and KCNA5 genes in samples from the RV Endocardium of controls (grey bars) and rTOF (black bars) hearts.
Transcript expression is normalized to the reference genes HPRT1 and GUSB. Two sided T-Test Statistical significance of n = 5 Controls and n = 7 rTOF hearts (*P <0.05, **P <0.01, ***P <0.001).
Table 1.
Fold change comparison between samples from the RV endocardium (rTOF (n = 7)r versus controls (n = 5)), RV epicardium (rTOF (n = 4) versus controls (n = 4)), LV epicardium (rTOF (n = 4) versus controls (n = 4)), and LV endorcadium (rTOF (n = 7) versus controls (n = 5)) based on microarray expression data of 14 genes.
Fig 7.
Relative expression (RT-qPCR) of genes in control (open bars) and Fallots (closed bars) pigs’ hearts and Fold change comparison between samples of the four localizations.
Transcript expression is normalized to the reference genes HPRT1 and GUSB. Two sided T-Test Statistical significance of n = 5 Controls and n = 7 rTOF hearts (*P <0.05, **P <0.01, ***P <0.001).