Neutrophil extracellular traps participate in the development of cancer-associated thrombosis in patients with gastric cancer

Figure 1 NETs are accumulated in samples of GC patients. A–C: Plasma levels of NET markers cf-DNA, citH3-DNA, and MPO-DNA in GC patients and healthy individuals were measured by ELISA. Healthy individuals, n = 13; stage I, n = 14; II, n = 15; III, n = 22; IV, n = 12; D-F: Comparison of cf-DNA, citH3-DNA and MPO-DNA in the plasma of patients with GC preoperatively and postoperatively by ELISA. n = 51; G and H: The rate of activated neutrophils in the circulating environment of GC patients and healthy individuals was measured by flow cytometry with APC-MPO and FITC-citH3 staining. Each group: n = 6; I and J: NET accumulation was detected by confocal microscopy with MPO and citH3 staining in paratumor and tumor samples from the same GC patient. Magnification 20×; scale bars: 50 μm. Red-citH3, Green-MPO and Blue-DAPI; K and L: Magnified (40×) part of MPO and citH3 colocation in paratumor and tumor samples from the same patient. Scale bars: 10 μm. Red-citH3, Green-MPO and Blue-DAPI; M: The percentage of area coverage of citH3 expression was defined as the rate of red area in total area and analyzed with ImageJ software; all values are the mean ± SD. ^aP < 0.05; ^bP < 0.01; ^cP < 0.001; ^dP < 0.0001. NET: Neutrophil extracellular trap; GC: Gastric cancer; cf-DNA: Cell-free DNA; MPO: Myeloperoxidase; citH3: Citrullinated histone H3.