Fig. 3

Neutrophil 1 cluster is associated with the formation of NETs in breast cancer. A tSNE and UMAP display the expression of NETs-related marker genes. B Volcano plot shows differentially expressed genes associated with NETs formation. C GO enrichment analysis of differentially expressed genes related to NETs formation. D KEGG enrichment analysis of differentially expressed genes related to NETs formation. E Differential genes of the Chemokine signaling pathway. F Differential genes of the IL-17 signaling pathway. G Differential genes of the Toll-like receptor signaling pathway. H ELISA detects the expression levels of NETs in the supernatant of neutrophils co-cultured with MCF7 and MDA-MB-231 cells. I ELISA detects the MPO-DNA expression levels in the supernatant of neutrophils co-cultured with MDA-MB-231 and HCC-1937 cells. J IF detects the expression levels of CitH3 and MPO in neutrophils co-cultured with MDA-MB-231 and HCC-1937 cells. K The staining of neutrophils with SYTOX GREEN after co-culture with MDA-MB-231 and HCC-1937 cells. L, M Assessment of tumor migration ability after co-culture with neutrophils or co-culture with neutrophils in the presence of a NETs inhibitor (DNase I). The data are shown as the means ± SDs; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, nonsignificant