Figure 3. In vivo binding of SPs-displaying phages to the endothelium in various regions of the mouse aorta.

The control phages, CLIRRTSIC- or CPRRSHPIC-displaying phages (2 × 10¹¹ pfu) were injected intravenously into C57BL/6 mice (n = 3) at 3 days post-LCA partial ligation, and allowed to circulate for 3 h. (a) Mouse aorta. The aorta was divided into four regions according to the vessel wall shear stress distribution: branch (BA, disturbed flow), greater curvature (GC, normal laminar flow), lesser curvature (LC, disturbed flow) and thoracic aorta (TA, normal laminar flow). (b) The aorta was removed and the level of phage binding evaluated by en face staining. Aortic tissues were fixed and bound phages were stained using an anti-M13 bacteriophage antibody (red); nuclei were stained with DAPI (blue). Attached phages were observed in GC, LC and TA regions of mouse aorta by confocal microscopy (red: phage, blue: nuclei, green: elastic lamina of vessel) (magnification, ×400; scale bars, 10 μm). Representative images are shown. Relative mean fluorescence intensity calculated using Image J for phage binding (red). *^{, #}Significant difference (*GC vs. LC; ^#TA vs. LC, p < 0.05).