Journal: Frontiers in Immunology

Article Title: The Presence of Anti-Lactoferrin Antibodies in a Subgroup of Eosinophilic Granulomatosis with Polyangiitis Patients and Their Possible Contribution to Enhancement of Neutrophil Extracellular Trap Formation

doi: 10.3389/fimmu.2016.00636

Figure Lengend Snippet: NET induction assay . Peripheral blood neutrophils obtained from healthy volunteers were seeded on chamber slides (1 × 10 5 /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.

Article Snippet: For in vitro assay, rabbit polyclonal anti-human Lf antibodies (CSB-PA00870EORb) (Cusabio Biotech, Hubei, China) and rabbit control IgG (Beckman Coulter, Tokyo, Japan) were employed.

Techniques: Incubation, Control, Positive Control, Software