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. 2022 Feb 3;41(5):e109386. doi: 10.15252/embj.2021109386

Figure 5. Bacterial signal peptides activate neutrophils and synergize with TLRs in Cxcl2 induction.

Figure 5

  • A
    Neutrophils from WT or Fpr‐deficient mice were allowed to migrate toward GBS signal peptide sequences (fPep1‐15, 1 µM), positive control peptides (f‐MIFL or WKYMVM, 1 µM) or vehicle (0.2% DMSO).
  • B
    Analysis of intracellular calcium flux in neutrophils stimulated with signal peptides as monitored by flow cytometry.
  • C, D
    Cxcl2 release in neutrophil cultures stimulated with combinations of HK‐bacteria, GBS fMet peptides, or TLR agonists. Neutrophils were exposed to a 50 µM concentration of Fpr agonists (fMIFL, WKYMVM or fPep8 and fPep10) for 1 h before the addition of the indicated TLR agonists, which included HK‐GBS (10 μg/ml), LPS (1 µg/ml), and RNA (1 µg/ml). In some cases, neutrophils were exposed to graded doses (10, 20, and 50 µM) of the indicated Fpr agonists alone.

Data information: Data are expressed as means + SD from three independent experiments, each conducted in duplicate. **P < 0.01, ***P < 0.001, as determined by unpaired t‐test.