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Fig. 5 VEGF-A-cGAS cascade-dependent biological network primarily targets cytoskeleton-related genes (A) Total number of DEGs in 3 dpf Tg(fli1a:eGFP);cgas−/− and Tg(fli1a:eGFP) larvae. (B) Heatmap of the top 200 most abundant transcripts in Tg(fli1a:eGFP);cgas−/− and Tg(fli1a:eGFP) larvae. (C) The top 50 up- (green) and downregulated (orange) DEGs in Tg(fli1a:eGFP);cgas−/− and Tg(fli1a:eGFP) larvae. (D) Network visualization of GO terms for vascular-related DEGs between Tg(fli1a:eGFP);cgas−/− and Tg(fli1a:eGFP) larvae. (E) GSEA enrichment plot. (F) KEGG analysis of overlapping genes. (G–J) The overlapped cGAS-targeted genes in different gene sets. (K–R) qRT-PCR confirmation of the overlapped genes in cGAS KO and vector control HUVECs with or without VEGF-A stimulation for 12 h (n = 3 replicates). (S) qRT-PCR assay showed most differentially expressed mRNAs are associated with cytoskeletal remodeling, cell adhesion/ECM remodeling, protein transport, and VEGF-A-VEGFR2 signaling pathways in cGAS KO and vector control groups of HUVECs (upper panel, n = 3 replicates). The potential targeted genes of the identified miRNAs were confirmed by RNA-seq analyses in Tg(fli1a:eGFP) and Tg(fli1a:eGFP);cgas−/− larvae (lower panel). Data are represented as means ± SEM. ANOVA in all analyses. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant. See also Table S1. Transcriptome analysis in, Table S2. Gene Ontology (GO) analysis showing the differentially expressed genes in, Table S3. List of genes that are both downregulated in, Table S4. KEGG analyzed the signaling pathways of overlapping genes that both downregulated in, Table S5. List of genes indicating that cGAS binding appeared onto cytoskeleton remodeling, cell adhesion ECM remodeling, vesicle-mediated protein transport, and overlapping genes, related to Figure 5.