Fig 1.
PCA clustering diagram and correlation heatmap of gene expression levels in RNA-seq data.
The x-label represents the variance contribution rate of principal component 1, and the y-label represents the variance contribution rate of principal component 2. The color of the heatmap indicates the correlation of gene expression levels in the sample, green represents low correlation and red represents high correlation. (A)-(B) GSE94019. (C)-(D) GSE102485.
Fig 2.
Volcano plots of two RNA-Seq data.
Red points represent up-regulated genes and blue points represented down-regulated genes. Gray points represent genes with no significant difference. (A) GSE94019. (B) GSE102485.
Fig 3.
The heatmap of the top 10 up-regulated and down-regulated genes in the RRA analysis.
Red indicates high expression of genes in patients with PDR and blue indicates low expression.
Fig 4.
The functional enrichment analysis for DEGs in GO terms and KEGG pathways.
(A) The bubble plot of top 10 significant GO terms in biological process (BP), cellular component (CC) and molecular function (MF). (B) The bubble plot of top 30 significant KEGG pathways.
Table 1.
Top five molecular functions in GO analysis.
Table 2.
Top five KEGG pathway enrichment analysis.
Fig 5.
t-distributed stochastic neighbour embedding (t-SNE) plot.
Purple represents Macrophages, orange represents Monocytes, red represents B cells, blue represents CD8 + T cells and green represents Fibroblasts.
Fig 6.
The expression level heatmaps of the top 10 DEGs.
(A) The top 10 DEGs in each cluster. (B) The top 10 DEGs in each cell type.
Fig 7.
The feature plots of each cell type marker genes.
Green represents high expression level and grey represents low expression level.
Fig 8.
43 ligand-receptor interactions and the PPI network of the interactions between fourteen proteins.
(A) 43 significant ligand-receptor interaction networks. (B) PPI network of interactions among CD4, CD44, COL1A1, COL1A2, COL4A1, COL4A2, COL6A2, GRN, ICAM1, LAMB1, NOTCH3, POSTN, SIGLEC1, and THY1. The color represents the degree of protein interactions in the PPI network, with darker color indicating the higher degree.
Fig 9.
Gene expression levels of CD4, CD44, COL1A1, COL1A2, COL4A1, COL4A2, COL6A2, GRN, ICAM1, LAMB1, NOTCH3, POSTN, SIGLEC1, and THY1 in each cell type.
(A) A dot plot shows the expression levels of these genes. The dot size and color represent the percentage of cells expressing the gene and p-values. (B) The violin plot shows the expression distribution of these genes.
Fig 10.
The Venn diagram shows 14 proteins that not only involved in significant ligand-receptor interactions in FVMs but DEGs-based PPI networks, three of which were associated with PDR.
Table 3.
The results of cellchat analysis including CD44, ICAM1 and POSTN.
Fig 11.
The heatmaps and regulation network of regulators.
(A) The heatmap of active regulators in each cell types. Red represents high regulatory activity and blue represents low activity. (B) The regulation network among 28 regulons and 3 key cellular communication genes. Diamond represents regulons and circle represents key cellular communication genes. (C) The heatmap of top five regulons in each cell types. Red represents high regulatory activity and blue represents low regulatory activity.
Fig 12.
Gene-gene co-expression heatmaps and protein interaction network.
(A) The co-expression heatmaps showing R values (the lower triangle) and P-values (the upper triangle), X targets means P-value > 0.05. Red represents positive correlation and blue represents negative correlation. (B) The MCL cluster analysis showing that POSTN, MYLK, ITGVA and ITGB5 are grouped into the same cluster, while CD44, ICAM1 and HIF1A are grouped into another cluster.
Fig 13.
Gene-pathway co-expression heatmap.
Red represents positive correlation and blue represents negative correlation. * Indicates p-value < 0.05, ** indicates p-value < 0.01.
Fig 14.
Results of validation using the GSE60436 dataset.
(A) A volcan plot showing POSTN and CD44 are significantly upregulated, but ICAM1 is not significantly changed. Red points represented up-regulated genes and blue points represented down-regulated genes, grey points represented genes with no significant difference. (B-D) Boxplots showing that POSTN and CD44 significantly upregulated, no significant change in ICAM1.