Microarray analysis of the time‑dependent expression profiles of long non‑coding RNAs in the progression of vein graft stenotic disease

Figures

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  Figure 1. Establishment of an external jugular vein carotid bypass rat model. (A) Exposure of the external jugular vein; (B) Exposure of the carotid artery; (C) The external jugular vein was anastomosed to the carotid artery; (D) The blood flow in the graft was patent.

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  Figure 2. Overview of significantly differentially expressed lncRNAs. (A) Heatmaps showing lncRNAs that were differentially expressed between groups. Scatter and volcano plots illustrate variations in lncRNA expression between different time points (0 vs. days 7, 14 and 28). (B) Chromosomal and (C) length distribution of differentially expressed lncRNAs. (D) Time series analysis revealed the temporal expression patterns of differentially expressed lncRNAs. Each box represents the time-dependent expression profile of a lncRNA. Upper value refers to the number of profiles; lower value indicates P-value of the profiles in each box. (E) Pie chart illustrating the classification of differentially expressed lncRNAs based on genomic location relative to neighboring or overlapping genes. (F) Time-dependent change in the number of differential expressed lncRNAs at three post-operative timepoints. lncRNAs, long non-coding RNAs.

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  Figure 3. ‘GO’ and ‘KEGG’ pathway enrichment analysis of lncRNA-associated protein-coding genes. (A) Significant BP, MF and CC terms in ‘GO’ enrichment analysis. (B) ‘KEGG’ enrichment analysis of significantly enriched pathways. P<0.05. GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; lncRNAs, long non-coding RNAs; BP, biological process; MF, molecular function; CC, cell component.

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  Figure 4. Biological processes associated with vascular remodeling and the interaction network between selected lncRNAs and associated protein-coding genes. (A) Time-dependent change in the number of differentially expressed lncRNAs predicted to be involved in ‘cell adhesion’, ‘migration’, ‘cell proliferation’, ‘MAP kinase activation’, ‘platelet activation’, ‘chemotaxis’, ‘WNT protein secretion’, ‘stimulation responses’ and ‘growth factor activity’. (B) Co-expression network between selected lncRNAs and associated protein-coding genes. Orange and blue circles indicate upregulated and downregulated lncRNAs, respectively. Green circles indicate protein-coding genes overlapped with mRNAs. lncRNA, long non-coding RNA.

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  Figure 5. Reverse transcription-quantitative PCR validation and Cell Counting Kit-8 assay. (A) Comparison of the expression levels of AF062402, BC091437 and BC166461 and associated protein-coding genes. (B) Expression of AF062402 in the Lentivirus-AF062402 group was significantly higher than that in the blank and Lentivirus-NC groups. Overexpression of AF062402 stimulated the proliferation of vascular smooth muscle cells. ****P<0.0001. NC, negative control; Mcam, melanoma cell adhesion molecule.