Dako a0082

All mice were sacrificed at 8 weeks. Explanted grafts were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned 5 μm thick. Sections were stained histologically with Carstair’s Method (Rowley Biochemical Inc. Danvers, MA) to distinguish fibrin and platelets, von Willebrand factor antibody (Dako A0082,1:1000, Agilent, Santa Clara, CA) for platelets, and Masson’s Trichrome for collagen fibers. For immunofluorescence staining, endothelial cells were stained using rabbit anti-CD31 primary antibody (1:50, ab28364, Abcam, Cambridge, UK) and smooth muscle cells were stained using mouse anti-a-SMA primary antibody (1:500, Agilent). Secondary fluorochrome conjugated antibodies were Alexa Flour 488 anti-rabbit immunoglobulin G secondary antibody (1:300, Invitrogen, Carlsbad, CA) and Alexa Fluor 647 anti-mouse immunoglobulin G secondary antibody (1:300, Invitrogen), respectively. Images were obtained under a fluorescence microscope (Zeiss AXIO Observer Z1, Oberkochen, Germany)