Figure 5.

Granulocyte colony-stimulating factor promotes NETosis in LNK-deficient mice. A, Wild-type (WT) or Lnk^–/– mice were intraperitoneally injected with vehicle or 2.5 μg recombinant human (rh) granulocyte colony-stimulating factor (G-CSF) for 15 minutes. p-STAT3 and citrullinated histone (H3Cit) levels in neutrophils isolated from WT or Lnk^–/– mice were determined and quantified by Western analysis (n=5). Unpaired t test. B, WT or Lnk^–/– neutrophils were stimulated with G-CSF for 3 hours. The cells were then stained with H3Cit (red) and DAPI and neutrophil extracellular traps (NETs) were quantified (n=5). C through E, WT or Lnk^–/– mice were treated with IP injection of vehicle or rhG-CSF (2.5 μg) for 3 days; neutrophils from vehicle or rhG-CSF–treated WT or Lnk^–/– mice were isolated and stimulated with WT platelets (C), Lnk^–/– platelets (D), or platelet-activating factor (E) for 4 hours. NETs were quantified (n=6); 2-way analysis of variance. Scale bar, 50 µm. Data are expressed as mean±SEM.