Fig. 2

GC cells activate neutrophils to release NETs, which promote the invasion and metastasis of GC cells. a Immunofluorescence staining was used to observe the difference in NET release from neutrophils between healthy volunteers and gastric cancer patients under the same conditions of stimulation. Neutrophils were stimulated with the supernatants of GES-1, MKN-45, AGS, and HGC-27 cells and characterized by cit-H3 (green) and MPO (red). b The percentage of NET cells released by NETs. c Cell-impermeable SYTOX Green staining was used to observe the difference in NETs released from healthy volunteers and gastric cancer patients under the same conditions of stimulation. Neutrophils and NETs (green). d The percentage of NET cells released by neutrophils. e, f The effect of NETs on the migration ability of the gastric cancer cell lines MKN-1 and HGC27 was detected using the scratch test. g The Transwell assay was used to detect the effect of NETs on the invasive ability of the gastric cancer cell lines MKN-1 and HGC27. h The number of cells passing through the compartment was counted using Image J software. Magnification, × 20; scale bars: 50 μm; red, cit-H3; green, MPO; blue, DAPI. All values are presented as means ± standard deviations. **p < 0.01; ***p < 0.001; ****p < 0.0001. GC, gastric cancer; GES-1, gastric mucosal epithelial cells; NET, neutrophil extracellular trap; MPO, myeloperoxidase; cit-H3, citrullinated histone H3