Fig. 2: CAF-derived factors drive NETosis systemically.

Quantification of A relative NET coverage and B relative number of bone marrow neutrophils taken from mice with pancreatic, lung or skin tumors. Quantification of the C relative NET coverage and D relative number of bone marrow neutrophils isolated from wild-type mice intravenously infused with pancreatic or lung FB or CAF CMed 24 h before analysing NETosis. E Quantification of relative NET coverage after stimulation with pancreatic CAF CMed with or without treatment with Cl-amidine in vitro. F Quantification of the relative NET coverage and G relative number of bone marrow neutrophils isolated from mice intravenously infused with lung CAF CMed with or without pre-treatment with Cl-amidine for 24 h. Data are mean ± SEM; *p < 0.05, **p < 0.01 and ***p < 0.001 using (A and F) paired t-test and C–E one-way ANOVA with a Dunnett post hoc test. Assays were performed on A–B n = 4 (in duplicate), n = 7 (in quadruplet) and n = 7 for pancreatic, lung and skin tumor-bearing mice respectively, C–D n = 8, 4 and 8 (Pancreatic for basal media, FB CMed and CAF CMed treatment, respectively. All performed in triplicate) and n = 9, 6 and 9 (Lung for basal media, FB CMed and CAF CMed treated, respectively. Performed in duplicate for basal media and CAF CMed and only a single time for FB CMed treated), E n = 3 (in triplicate) and F–G n = 8 (in triplicate) independent experiments.