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. 2016 Apr 14;7:137. doi: 10.3389/fimmu.2016.00137

Figure 6.

Figure 6

Anti-CFP antibody blocks activation of complement AP to form C5b–9 on PMA-induced NETs. Neutrophils were activated with PMA (20 nM) to induce NET formation. Plasma with RPMI buffer was used to activate all complement pathways, AP buffer to only activate the alternative pathway. A monoclonal anti-properdin antibody was used to inhibit AP activation. A buffer exchange was performed followed by addition of 20% (v/v) plasma:RPMI1640 + 10 mM Hepes [far left column (A,E,I)], 20% (v/v) plasma:RPMI1640 + 10 mM Hepes + mAb properdin [left column (B,F,J)], 20% (v/v) plasma:AP buffer [right column (C,G,K)], 20% (v/v) plasma:AP buffer + mAb properdin [far right column (D,H,L)]. (A–D) show DAPI, (E–H) C5b–9 deposition, and (I–L) merged images. Images were taken with 60×/1.35 oil immersion objective. (I–K) with 2× magnified insets of representative areas. Scale bar, 9.00 μm.