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. Author manuscript; available in PMC: 2017 Aug 10.
Published in final edited form as: Sci Transl Med. 2016 Oct 19;8(361):361ra138. doi: 10.1126/scitranslmed.aag1711

Figure 4. Cancer cells induce NET formation through G-CSF, and neutrophil-stimulated invasion requires NADPH oxidase and PAD4 activity.

Figure 4

(A) Blocking anti-G-CSF antibodies (1.6 μg/mL) decreased 4T1-induced NET extension (mean ± SEM, neutrophils with vehicle or anti-G-CSF: n=3; neutrophils with 4T1 cells and vehicle or anti-G-CSF: n=5, t-test).

(B) The NADPH oxidase inhibitor apocynin (10 μM) inhibited NET formation (mean ± SEM, neutrophils and vehicle or NAPDH oxidase inhibitor: n=3, neutrophils with cancer cells and vehicle or apocynin: n=5, t-test).

(C) Apocynin (10 μM) inhibited neutrophil-stimulated invasion (mean ± SEM, 4T1 cells only or 4T1 cells with neutrophils and vehicle or apocynin: n=4; 4T1 cells only and vehicle or apocynin in 10% FCS: n=1, t-test).

(D) PAD4 inhibition (200 μM Cl-amidine) reduced cancer cell-induced NET formation (mean ± SEM; neutrophils and vehicle or PAD4 inhibitor: n=6; neutrophils with 4T1 cells and vehicle or PAD4 inhibitor: n=4, t-test).

(E) PAD4 inhibition (200 μM Cl-amidine) blocked neutrophil-stimulated cancer cell invasion (mean ± SEM; n=3, t-test).