Role of Macrophage‐Related Genes GAS7 and ZEB2 in Acute Myocardial Infarction Pathogenesis

Figures

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  Figure 1. Single‐cell clustering of AMI. (A–B) Construction of single‐cell maps of AMI and control samples. (C) Bubble chart showing the expression levels of marker genes for each cell type. (D) Differences in cell proportions between AMI and control samples.

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  Figure 2. Analysis of communication between cells in AMI. (A–B) Comparison of the number and intensity of cell communication events between AMI and the control group. (C–D) Network diagram of the number and intensity of intercellular communication in the AMI group, with red arrows indicating strong communication and thicker arrows indicating higher weight. (E–F) Communication network diagram between cell types in the control group. (G) Comparison of signaling pathways with significant changes in relative information flow. (H–I) Bubble plot of ligand–receptor pairs in AMI and control samples, with dot size indicating p value and color indicating significance.

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  Figure 3. GSEA analysis results: (A) GSEA analysis of macrophages, (B) cardiomyocytes, (C) endothelial cells, and (D) fibroblasts in AMI.

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  Figure 4. hdWGCNA screening of macrophage‐related modules in AMI. (A) Soft threshold screening, with the optimal soft threshold in the figure being 6. (B) Coexpression network tree, with the upper part representing the gene tree, where each branch corresponds to a gene, and the lower part representing the modules associated with each gene. (C) Module partitioning, with the vertical axis representing kME values, indicating the connectivity of each gene based on feature genes and the right side showing the hub genes of each module.

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  Figure 5. Expression of genes within modules in different cells. (A) Expression levels of genes within modules in different cells, with red representing high expression, purple representing low expression, and circle size representing cell proportion. (B) Violin plots of module feature value expression distributions in each cell for coexpression Modules M5 and M8. (C) Coexpression network of hub genes in coexpression Modules M5 and M8.

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  Figure 6. Transcription factor identification in macrophages. (A) Distribution of key regulatory factor activity in AMI and donor samples. (B) GO functional enrichment analysis of the regulon target gene set. (C) Transcription factor–target gene regulatory network diagram, with orange triangles representing transcription factors and green circles representing target genes.

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  Figure 7. WGCNA analysis results of AMI‐related genes. (A) Soft threshold selection was used to determine the scale‐free nature of the network. (B) The gene clustering dendrogram shows the distribution of different modules. (C) Heat map showing the correlation between different gene modules and AMI and the control group. (D) Number of genes in each module. (E) Scatter plot showing the correlation between gene module membership and gene significance in the brown module. (F) Functional enrichment analysis of genes in the brown module.

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  Figure 8. Molecular docking of biomarkers. (A) Venn diagram showing the intersection of key candidate genes screened by three methods: hdWGCNA, TF regulatory network, and macrophage‐specific WGCNA. (B) ROC curves for key genes. (C) Expression of key genes in GSE66360. (D) Molecular docking results of GAS7 and sulforaphane. Blue molecules represent receptor proteins, orange molecules represent drug small molecules, and cyan represents amino acids. Hydrogen bonds between receptor proteins and drug small molecules are indicated by yellow dashed lines, with the numbers above representing hydrogen bond lengths.

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  Figure 9. Validation of key genes in an OGD‐induced injury model. Relative mRNA expression levels of FOS, ETV6, GAS7, and ZEB2 in control and OGD‐treated cells measured by qRT–PCR (n = 3). Data are presented as mean ± SD. ∗∗∗ p < 0.001 versus control.