Fig 2: CNTN2 is downregulated in osteogenically induced hVICs, which is associated with osteogenic differentiation. (A) Volcano plot showing the upregulated and downregulated genes in osteogenically induced hVICs (CNTN2: logFC=−4.286, adjusted p-value < 0.001). (B) GO analysis reveals differentially expressed genes enriched in biological processes. (C) KEGG analysis of differentially expressed genes enriched in pathways. The KEGG imagery used in this figure was obtained from the KEGG database, and we acknowledge the permission provided by the Kanehisa laboratory. This figure is cited in accordance with KEGG citation guidelines (www.kegg.jp/kegg/kegg1.html)20. (D) Heatmap showing that the expression of CNTN2 is significantly downregulated in hVICs induced by OM. (E) Alizarin Red staining demonstrated the osteogenic differentiation of hVICs and relative calcium deposits induced by OM, n = 6. Western blotting revealed the expression levels of CNTN2, ALP, and RUNX2 in the control (con) and OM groups, n = 6. GO, gene ontology; KEGG, kyoto encyclopedia of genes and genomes, CNTN2, Contactin 2; OM, osteogenic media; hVICs, human primary aortic valve interstitial cells; ALP, alkaline phosphatase; RUNX2 runt-related transcription factor 2.