Figure 9.
ALF results in the formation of NETs in the liver under regulation of NE. (A) Immunofluorescence of H3Cit (red), NE (green), and DAPI (blue) in liver sections showing NE as a component of NETs in mouse liver with ALF (scale bar: 5 μm). (B) The formation of NETs was detected by immunofluorescence of H3Cit (red), NE (green), and DAPI (blue) in mouse primary neutrophils with ex vivo stimulation by LPS (200 μg mL-1) for 3 hours (scale bar: 1 μm). Immunofluorescence of H3Cit (red), MPO (green), and DAPI (blue) in primary neutrophils isolated from NE KO mice and WT control mice with ex vivo stimulation by LPS (C, scale bar: 5 μm) to identify the formation of NETs and (D) semiquantification under 40× microscopic fields. Representative images of immunofluorescence of H3Cit, MPO, and DAPI in liver sections from 10-week-old male NE KO mice and WT control mice treatment with (E, scale bar: 5 μm) D-GalN/LPS or PBS as vehicle and (F) semiquantification of the number of NETs under 40× microscopic fields, indicating the impaired capacity for NET formation in NE KO mice under ALF conditions. Ex vivo data were analyzed according to 3 independent experiments. Data in animal studies are expressed as means ± SEM. n = 5. ∗∗P < .01.