Transcriptome sequencing and lncRNA-miRNA-mRNA network construction in cardiac fibrosis and heart failure

Figures & data

Figure 1. Volcano map, heatmap and venn diagram analysis of DEmRNAs in the GC/GD and GH/GD groups.

A: Volcano map of DEmRNAs in the GC/GD group; B: Heatmap of DEmRNAs in the GC/GD group; C: Volcano map of DEmRNAs in the GH/GD group; D: Heatmap of DEmRNAs in the GH/GD group; E: Venn diagram of up-regulated DEmRNAs; F: Venn diagram of down-regulated DEmRNAs. In the volcano map, horizontal axis is the value of Log2 logarithmic conversion, and the vertical axis is p-value after logarithmic conversion of -Log10. Heatmap shows result of a two-way hierarchical clustering of DEmRNAs (row) and samples (column). DEmRNAs clustering tree is shown on the left. Red and green represent up-regulated and down-regulated DEmRNAs, respectively. GD, GC, and GH represent the normal control group, the myocardial fibrosis group and the heart failure group, respectively.

Figure 2. Volcano map, heatmap and Venn diagram analysis of DElncRNAs in the GC/GD and GH/GD groups.

A: Volcano map of DElncRNAs in the GC/GD group; B: Heatmap of DElncRNAs in the GC/GD group; C: Volcano map of DElncRNAs in the GH/GD group; D: Heatmap of DElncRNAs in the GH/GD group; E: Venn diagram of up-regulated DElncRNAs; F: Venn diagram of down-regulated DElncRNAs. In the volcano map, horizontal axis was the value of Log2 logarithmic conversion, and the vertical axis is p-value after logarithmic conversion of -Log10. Heatmap shows result of a two-way hierarchical clustering of DElncRNAs (row) and samples (column). DElncRNAs clustering tree is shown on the left. Red and green represent up-regulated and down-regulated DElncRNAs, respectively. GD, GC, and GH represent the normal control group, the myocardial fibrosis group and the heart failure group, respectively.

Figure 3. Volcano map, heatmap and Venn diagram analysis of DEmiRNAs in the GC/GD and GH/GD groups.

A: Volcano map of DEmiRNAs in the GC/GD group; B: Heatmap of DEmiRNAs in the GC/GD group; C: Volcano map of DEmiRNAs in the GH/GD group; D: Heatmap of DEmiRNAs in the GH/GD group; E: Venn diagram of up-regulated DEmiRNAs; F: Venn diagram of down-regulated DEmiRNAs. In the volcano map, horizontal axis is the value of Log2 logarithmic conversion, and the vertical axis is p-value after logarithmic conversion of -Log10. Heatmap shows result of a two-way hierarchical clustering of DEmRNAs (row) and samples (column). DEmRNAs clustering tree is shown on the left. Red and green represent up-regulated and down-regulated DEmiRNAs, respectively. GD, GC, and GH represent the normal control group, the myocardial fibrosis group and the heart failure group, respectively.

Figure 4. Significantlly enriched GO terms and KEGG pathways of common DEmRNAs.

A: Biological process (BP); B: Cell composition (CC); C: Molecular function (MF); D: Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The x-axis shows counts of DEmRNAs enriched in GO terms or KEGG pathways, and the y-axis shows GO terms or KEGG pathways. Red represents up-regulated DEmRNAs and blue represents down-regulated DEmRNAs.

Table 1. DEmiRNA-DEmRNA targeted relationship

DEmiRNA		DEmRNA
Name	Up/Down	Name	ID	Up/Down
hsa-miR-144-3p	Down	CCNE2	ENSG00000175305	Up
		C9orf72	ENSG00000147894	Up

DEmiRNA: Differentially expressed miRNAs; DEmRNA: Differentially expressed mRNAs; Up: Gene expression levels were up-regulation; Down: Gene expression levels were down-regulation.

Figure 5. DEmiRNA-DElncRNA targeting network diagram (a) and DElncRNA-DEmiRNA-DEmRNA interaction network (b).

Square, circle, V-shape, red and green represents DEmiRNA, DEmRNA, DElncRNA, up-regulation, and down-regulation, respectively.

Table 2. Primer sequence in the RT-PCR

Primer name	Primer sequence (5’ to 3’)
GAPDH-F (Internal reference)	5-CTGGGCTACACTGAGCACC-3
GAPDH-R (Internal reference)	5-AAGTGGTCGTTGAGGGCAATG-3
ACTB-F (Internal reference)	5-CATGTACGTTGCTATCCAGGC-3
ACTB-R (Internal reference)	5-CTCCTTAATGTCACGCACGAT-3
CCNE2-F	5-AGGAATTGTTGGCCACCTGT-3
CCNE2-R	5-TCCCCAGCTTAAATCAGGCA-3
C9orf72-F	5-TGGGACATGACCTGGTTGC-3
C9orf72-R	5-TCAACGCGGCCAGATAGAC-3
LEF1-AS1-F	5-AGCCGAATTTCCTTAGCCGT-3
LEF1-AS1-R	5-CCACACGTGTTGTGTCAACG-3
MLK7-AS1-F	5-CCTGCAGCACGTTTCCATG-3
MLK7-AS1-R	5-GCCAAATCCAGACCCACCT-3
hsa-miR-144-3p-F	5-TACAGTATAGATGATGTACT-3

Figure 6. Expression validation of CCNE2, C9orf72, LEF1-AS1, MLK7-AS1, and hsa-miR-144-3p in blood samples by RT-PCR.

GD, GC, and GH represent the normal control group, the myocardial fibrosis group, and the heart failure group, respectively. * represents p < 0.05.

Figure 7. Expression validation and diagnostic analysis of CCNE2 and hsa-miR-144-3p.

A: Expression validation of CCNE2; B: Expression validation of hsa-miR-144-3p; C: ROC curve of CCNE2; D: ROC curve of hsa-miR-144-3p.