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. 2012 Jul 31;31(17):3607–3619. doi: 10.1038/emboj.2012.212

Figure 2.

Figure 2

Staphopain A inhibits CXCR2-mediated calcium mobilization of neutrophils. Fluo-3-labelled neutrophils were preincubated with buffer or 0.5 μM Staphopain A with or without 1 μM Staphostatin A, for 15 min at 37°C. After washing, cells were stimulated with different concentrations of CXCL1 (A), CXCL7 (B), CXCL8 (C), fMLF (D), C5a (E) or a fixed concentration (3 × 10−9 M) of CXCL2, CXCL3, CXCL5 and CXCL6 (F). To determine the IC50 (50% of inhibition), Fluo-3-labelled neutrophils were preincubated with different concentrations of Staphopain A for 15 min at 37°C and subsequently stimulated with 3 × 10−9 M CXCL1 (G) or 1 × 10−8 M CXCL7 (H). The IC50 for CXCL1 and CXCL7 was calculated with the formulas y=−4E+10−7x+4.1314 and y=−3E+10−7x+4.0248, respectively. All figures represent the mean±s.e. of three separate experiments using different donors. The relative calcium mobilization was calculated by dividing the fluorescence after stimulation by the baseline fluorescence. *P<0.05 versus buffer; **P<0.01 for Staphopain A versus buffer (two-tailed Student’s t-test).