Figure 2. NET demonstration by immunostaining.
2 x 105 human neutrophils (A) or murine neutrophils (B) were activated by 100 ng/ml PMA for 3 h in wells of glass chamber slide. Resting neutrophils with DMSO served as negative control. Slides were fixed and labelled with anti-DNA/Histone H1 antibody followed by AF555-conjugated anti-mouse IgG (red) and nuclei were counterstained with DAPI (blue) (steps B1 and B2). C. Immunostaining of human neutrophil marker CD15 (green) and NET marker DNA/Histone H1 (red) on paraffin-embedded skin section from human vasculitis patients. D. Immunostaining of the murine neutrophil marker Ly6G (green) and NET marker DNA/Histone H1 (red) on paraffin-embedded skin section from mice with induced immune complex-mediated vasculitis (step B3). Scale bars = 100 µm.