Fig 2: FOXS1 expression is positively correlated with angiogenesis and CXCL8 expression in CRC. (A) Correlation analysis of FOXS1 and CD31, CD34, CD105, or CD146 in GSE39582 and TCGA-CRC. (B) Correlation analysis of FOXS1 and CXCL8 in GSE39582 and TCGA-COAD (r = 0.110, p < 0.01; r = 0.237, p < 0.001). (C) The expressions of FOXS1, CXCL8, and CD31 protein in specimens, including 39 CRC tissue specimens, were examined by IHC. Representative IHC images (left) and correlation analysis (right) of FOXS1 and CXCL8 expression or microvessel count (indicated by CD31-positive cells) in per field are shown. Scale bars: 50 µm. ***p < 0.001.

Fig 3: Molecular docking between CXCL8 and five pivotal ingredients, including quercetin (A), beta-sitosterol (B), kaempferol (C), palmidin A (D), and candletoxin A (E).

Fig 4: FOXS1 promotes EMT and angiogenesis by upregulating CXCL8. (A) Venn diagram analysis across three groups of genes (left). (i) Angiogenesis signature genes (n = 48); (ii) differentially expressed tumor versus normal in GSE18105 (n = 794); (iii) secreted protein genes (n = 1,708). The FOXS1 motif (orange boxes) was searched on the promoter loci of the CXCL8 gene and VEGFA gene in the JASPAR database. (B) Real-time PCR analysis of CXCL8 expression in CRC cell lines. (C) Real-time PCR analysis of the expression of epithelial–mesenchymal transition markers in CRC cell lines. (D) Transwell migration assay and transwell invasion assay were performed to determine the effects of CXCL8 depletion on the migratory and invasive abilities of FOXS1-overexpressed CRC cells. Scale bars: 50 µm. (E) HUVEC tube formation after stimulation with the indicated conditional medium from FOXS1-overexpressed CRC cells with or without CXCL8 depletion. Scale bar: 100 µm. ***p < 0.001.

Fig 5: Screening of HUB genes. (A) PPI network of WMW against SDA performed by STRING database with the parameters of the organism for “Homo sapiens” and the minimum required interaction score= 0.9. (B) 35 HUB genes network performed by Cytoscape yFiles Radial Layout. Low degree values showed large sizes and bright colors, and small sizes and dark colors for high degree values on the contrary. (C) The 10 core targets network, including JUN, MAPK1, IL6, TP53, RELA, HSP90AA1, EGFR, AKT1, CXCL8, and TNF.