Fig. 2
C5a promoted arterial thrombosis by triggering neutrophils to release NETs. A, B H&E staining of thrombus, cross-sections (A) and longitudinal sections (B). Scale bar = 100 µm. C Quantification of the thrombus size in cross-sections by H&E staining (n = 5 each). D Thrombus weight (n = 5 each). E The blood flow velocity in the LICA was dramatically decreased in the FeCl3-induced thrombus mice compared with the sham mice, and this change was reversed by PMX53. F Quantification of the blood flow velocity (n = 4 each). G Immunofluorescence staining of cross-sections of the LCCA for CitH3 (red), Ly6g (green) and DAPI (blue) 6 h after FeCl3 exposure. Thirty minutes before FeCl3 exposure, PMX53 was administered by intraperitoneal injection. Scale bar = 75 µm. H Quantification of NET formation capacity shown as the ratio of NETs/neutrophil in LCCA cross-sections subjected to immunofluorescence staining (NaCl group = 3, PMX53 group = 4). I Representative images of immunofluorescence stainings of NET formation for DNA (SYTOX green), CitH3 (red), Ly6g(blue) in vitro after stimulation of C5a, and NET formation was attenuated by PMX53. Scale bar = 75 µm. J Quantification of NET formation capacity in vitro shown as the percentage of NET release, which was assessed by immunofluorescence staining (n = 3 each). (K) NETosis was measured using a plate reader assay. NET release is expressed as percentage of total DNA (n = 3 each). Data are presented as mean ± SD