Fig. 5. NETs-GC clusters facilitate GC cells extravasation and survival at metastatic sites.

A–D Schematic experiment design for NETs-GC clusters in a specific transwell assay (A). The mixture of MKN-45 cells and neutrophils from indicated groups were subjected to upper chamber. Then the penetrated NETs-GC clusters in upper chamber membrane were counted under a microscopy (B) and that in the lower chamber medium were analyzed by a SEM, in which green arrows point to extracellular meshes of NETs, white arrows point to MKN-45 cells and red arrows point to neutrophils (C); D The mixture of MKN-45 cells and neutrophils from AIC group were subjected to transwell assays with or without LY 2157299 added to the lower chamber medium. Then penetrated cells in the upper chamber membrane were counted under a microscopy; E Representative immunofluorescence co-staining images of DNA, Cit-H3, and MPO to assess NETs and GC cells extravasation in the liver of LM nude mice (left panel) or implantation in the peritoneum of PM nude mice (right panel) at post-modeling day 1 (n = 3 per group); F Representative immunofluorescence co-staining images of DNA, Cit-H3 and MPO to assess NETs and GC cells extravasation in the liver of LM nude mice (left panel) or implantation in the peritoneum of PM nude mice (right panel) at post-modeling day 5 (n = 3 per group). Data represent the mean ± S.D. in B, C, D (n = 3 biologically independent experiments), one-way ANOVA with Tukey test was used in B, C. paired Student’s t-tests were used in D. Source data are provided as a Source Data file.