Figure 3.
NET induction assay. Peripheral blood neutrophils obtained from healthy volunteers were seeded on chamber slides (1 × 105/mL), incubated for 15 min at 37°C, and then exposed to 0 or 10 nM phorbol myristate acetate (PMA) combined with 1 µg/mL aLf (CSB-PA00870EORb) or control rabbit IgG. After incubation for 3 h at 37°C, the samples were fixed with 4% paraformaldehyde followed by mounting with a solution containing DAPI. For positive control, the neutrophils were exposed to 100 nM PMA for 3 h at 37°C. (A) The representative photomicrographs are shown (original magnification: ×200). (B) NET area was represented by DAPI-positive area, which was calculated using Image J software. Data were presented as mean ± SD values of relative NET induction in which the value of the positive control (PMA 100 nM) was set as 1. Experiments were repeated five times. **p < 0.01.