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. 2017 Feb 9;8:60. doi: 10.3389/fimmu.2017.00060

Figure 5.

Figure 5

Toll-like receptors (TLRs) were involved in Staphylococcus aureus (S. aureus)-induced but not phorbol 12-myristate 13-acetate (PMA)-induced neutrophil extracellular traps (NETs) formation. Neutrophils were pretreated with TLRs agonist or antagonist, followed by PMA or S. aureus stimulation. NETs formation was quantified by Quant-iT PicoGreen double-stranded deoxyribonucleic acid assay kit. (A) None of TLR2 agonist (HKLM), TLR4 agonist (LPS), TLR5 agonist (FSL-ST), and TLR6 (FSL-1) agonist could induce NETs formation. (B) Treatment with TLR2 agonist (HKLM) and TLR4 agonists (LPS) significantly enhanced NETosis, and blocking TLR2 and TLR4 with neutralizing antibodies significantly reduced S. aureus-induced NETs formation. None of TLR5 agonist (FSL-ST), TLR6 agonist (FSL-1), and TLR5 and TLR6 neutralizing antibodies was involved in S. aureus-induced NETs formation. (C) TLRs were not involved in PMA-induced NETs formation. The assay was repeated for three times, each case in five wells, error bars represent SEM. Compared to S. aureus or PMA stimulation, *p < 0.05, ns = p > 0.05 by Student’s t-test.