Mouse anti collagen 1

Immunofluorescence staining was performed as previously described [5 (link)]. Smooth muscle cells were stained using mouse anti-SM-MHC (Abcam, UK) primary antibodies. Endothelial cell staining was performed using rabbit anti-vWF (Abcam, USA) primary antibody. The vascular precursor cell was stained using rabbit anti-Sca-1 (Millipore, Germany). For elastin and collagen staining, slides were incubated with rabbit anti-Elastin (Abcam, UK), mouse anti-Collagen I (Abcam, UK) and mouse anti-Collagen III (Abcam, UK). To observe inflammatory cells in the explanted grafts, mouse anti-CD68 (Abcam, UK), rabbit anti-iNOS (Abcam, UK) and rabbit anti-CD206 (Abcam, UK) were used as primary antibodies. After overnight incubated at 4 °C, slides were washed twice with PBS solution and incubated with the respective fluorescein isothiocyanate-conjugated secondary antibodies for 60 min at 37 °C. The samples were observed using an Olympus Flu view 1000 confocal microscope (Japan). Tissue slides pretreated without a primary antibody were used as negative controls. Cell populations were determined based on cell counts from each image on six different parts (12, 2, 4, 6, 8, and 10 o'clock positions). Data were collected from three different samples in each group. Details of the primary antibodies are listed in Table S1.