Fig. 6
From: Neutrophil extracellular trap induced by HMGB1 exacerbates damages in the ischemic brain
Blocking HMGB1 suppressed NETosed neutrophil-induced neuronal cell death. a Schematic diagram of the procedure for blocking HMGB1 in the co-culture of NETosed neutrophils and primary cortical neurons. b-c PMNs-BM (5 × 105/well) were treated with NCM (1.6 × 106/4 well) for 3 h and then cultured in fresh media for another 3, 6, 12, or 18 h and HMGB1 levels were examined in cell lysate and culture media by immunoblotting (b). NET formation was visualized after 6 h of NCM treatment by double immunofluorescent staining using anti-HMGB1 antibody and DAPI (c). d-e NCM-treated PMNs were co-cultured with naïve primary cortical culture (4 × 105/well) for 18 h in the presence or absence of anti-HMGB1 antibody (1 μg/ml) or HMGB1 A box (50 ng/ml) and the numbers of PI-positive cells were counted (250 μm × 250 μm). Scale bars in C represent 20 μm and those in E represent 100 μm. Results are presented as means±SEMs. **p < 0.0