The induction of adhesion molecules on ECs by sE-selectin. (A) RT-PCR analysis of ICAM-1 and VCAM-1 expression on ECs after stimulation with sE-selectin (sE-sel). (B) FACS analysis of ICAM-1 and VCAM-1 protein expression on ECs. sE-selectin increased both mRNA and protein expressions of ICAM-1 remarkably and VCAM-1 mildly in ECs. VEGF was used as positive control. (C,D) Immunofluorescent images of ICAM-1 and VCAM-1 in nonischemic (C) and ischemic hindlimb (D). The expression of ICAM-1 (red) on endothelium (green) was induced by injecting sE-selectin into nonischemic muscle (C, scale bar, 200 μm). Overexpressed ICAM-1 by ischemia was suppressed when E-selectin was neutralized by injecting anti–E-selectin antibody into ischemic muscle (D, scale bar, 200 μm). The expression of VCAM-1 was low in all conditions. DAPI (blue) staining was performed to visualize the nucleus.