Fig 1. Pancreatic cancer cells AsPC-1 induce neutrophil extracellular traps.

(A) AsPC-1 (5 × 10⁴ cells/mL) were incubated with whole blood for 30 min, 1 h, or 2 h at 37°C. Control represents whole blood without AsPC-1. The histone–DNA complex levels were measured in the supernatants. A significant difference was observed among the groups in histone-DNA complex level (NET formation) during incubation time. Statistical analysis was performed by two-way ANOVA followed by Bonferroni’s post-hoc test. ^*P < 0.05 versus control group. (B) AsPC-1 (5 × 10⁴ or 20 × 10⁴ cells/mL) were incubated with whole blood for 2 h. Phorbol 12-myristate-13-acetate (PMA, 25 nM) was used as a positive control of NET formation. Statistical analysis was performed by one-way ANOVA followed by Bonferroni’s post-hoc test. *P < 0.05 versus whole blood without AsPC-1. (C) AsPC-1 cells were incubated with isolated neutrophils for 2 h and the histone–DNA complex level was measured in the supernatants. (D) Conditioned medium (CM) harvested from AsPC-1 culture was incubated with whole blood for 2h and the histone-DNA complex level was measured in the supernatants. Charcoal was added to CM to remove lipids and heat treatment was used to degrade proteins. Data are expressed as mean ± SEM of 3 experiments. ^*P < 0.05 versus control; **P < 0.05 versus CM. Abbreviations: AU, arbitrary units.