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After 1?h incubation at room temperature, the contents of the tubes were mixed by inverting them 3�C4 times. After centrifugation for 10?min at 1750?g, hemolysis was determined in the supernatant by measuring the absorbance of released hemoglobin at http://www.selleckchem.com/products/Adriamycin.html 412?nm compared to the references. Cryostat sections of the pig liver xenografts were fixed in acetone and incubated with the following primary antibodies overnight �C mouse anti-porcine P-selectin (clone 12C5) and CD106 (10.2C6) (generous gifts from Professor D.O. Haskard, Imperial College London, UK); custom rabbit anti-porcine TF raised against a synthetic peptide comprising the sequence IMRNVKETYV present in the porcine TF protein (NCBI reference sequence NP_998950.1); mouse anti-porcine E-selectin (clone 1.2B6; Sigma); mouse anti-human vWF (clone F8/86; DAKO, Carpinteria, CA, USA); mouse http://www.selleckchem.com/products/obeticholic-acid.html anti-primate CD45 (clone 5H9; BD); mouse anti-human CD42a (clone fmc25; AbDSerotec, Raleigh, NC, USA); sheep anti-human TF (Affinity Biologicals); sheep anti-human fibrin (clone SAFNE; Affinity Biologicals); mouse anti-porcine CD31 (clone APG311; Antigenix America, Huntington Station, NY, USA) [17,18]; anti-human CD41 (clone ab63983; Abcam, Cambridge, MA, USA); rabbit anti-human IgG (DAKO), rabbit anti-human IgM (DAKO); rabbit anti-human C3 (DAKO); mouse anti-human C5-9 (DAKO); mouse anti-human CD68 (DAKO); mouse anti-human CD20 (DAKO); rabbit anti-human CD3 (DAKO). After washing, the sections were incubated with appropriate secondary antibodies for 1?h [CyChrome 2 anti-sheep IgG, CyChrome 3 anti-mouse IgG, CyChrome 5 anti-rabbit IgG (Jackson ImmunoResearch, West Grove, PA, USA)]. Nuclei were stained with DAPI (4,6-diamidino-2-phenylindole; http://www.selleck.cn/products/sch772984.html Molecular Probes, Eugene, OR, USA). After paraformaldehyde-fixation, the tissues were prepared with poly-L-lysine-coated slides. Images were viewed through a Nikon E-800 microscope (Melville City, NY, USA). Liver tissue was fixed with 2.5% glutaraldehyde in PBS. Transmission electron microscopy was performed, as previously described [19]. Data are presented as mean?��?SEM. Significance of the difference between two groups was determined by paired Student��s t test. Values of P?