Fig. 6

NET-DNA promoted CCA cell lines synthesized VEGF-a via ITGAV/ NFκB axis and induced angiogenesis. A Representative images of CD34 staining showed that increased angiogenesis in WT mice group compared to the PAD4 KO mice group. Scale bar=50μm. B Elevated angiogenesis_score in high NETs_score group compared to the NETs_low score group. C Positive correlation between Cit-H3 and CD34 expression in liver sections from patients with CCA. D Correlation of protein expression of CD34 and CitH3 (n=37, p=0.0077, Spearman’s coefficient r=0.401). E Chip-qPCR assay showed NFκB can bind to the VEGF-A promoter regulatory region. F VEGF-A ELISA assay showed that NET-DNA stimulated ITGAV/NFκB pathway to induce VEGF-A biogenesis. G The conditioned medium from CCA cell lines was employed in a tube formation assay, revealing that NET-DNA promotes angiogenesis through the ITGAV/NFκB pathway. Scale bar=20μm. Data are means ± SD of three independent experiments. **P<0.01, ***P<0.001