Fig 2: Collagen‐binding activities of glycosidase‐treated VNs. Collagen type I was immobilized on wells of a 96‐well plate. The purified glycosidase‐treated VNs were added to the wells and reacted with the collagen in 10 mm Tris/HCl saline (pH 7.5). The VNs bound to collagen were detected by rabbit anti‐VN IgGs (1/25 000, LSL‐LB‐2096; Cosmo Bio Co., Ltd) and goat anti‐rabbit IgGs Fc‐HRP (1/10 000, AP156P; Millipore Corp.). Binding studies were carried by ELISA as described in Materials and methods. The absorbance of collagen‐bound VNs was corrected using the reactivity of each VN to anti‐VN IgGs. C, control VN incubated without enzyme (●); F, VN treated with fucosidase (▲); N, VN treated with neuraminidase (♦); F+N, VN treated with both fucosidase and neuraminidase (□); NG, VN treated with N‐glycosidaseF (○). (A) hVN, (B) pVN. Data are presented as mean ± SD.

Fig 3: Collagen‐binding activities of glycosidase‐treated VNs. Collagen type I was immobilized on wells of a 96‐well plate. The purified glycosidase‐treated VNs were added to the wells and reacted with the collagen in 10 mm Tris/HCl saline (pH 7.5). The VNs bound to collagen were detected by rabbit anti‐VN IgGs (1/25 000, LSL‐LB‐2096; Cosmo Bio Co., Ltd) and goat anti‐rabbit IgGs Fc‐HRP (1/10 000, AP156P; Millipore Corp.). Binding studies were carried by ELISA as described in Materials and methods. The absorbance of collagen‐bound VNs was corrected using the reactivity of each VN to anti‐VN IgGs. C, control VN incubated without enzyme (●); F, VN treated with fucosidase (▲); N, VN treated with neuraminidase (♦); F+N, VN treated with both fucosidase and neuraminidase (□); NG, VN treated with N‐glycosidaseF (○). (A) hVN, (B) pVN. Data are presented as mean ± SD.

Fig 4: Collagen‐binding activities of glycosidase‐treated VNs. Collagen type I was immobilized on wells of a 96‐well plate. The purified glycosidase‐treated VNs were added to the wells and reacted with the collagen in 10 mm Tris/HCl saline (pH 7.5). The VNs bound to collagen were detected by rabbit anti‐VN IgGs (1/25 000, LSL‐LB‐2096; Cosmo Bio Co., Ltd) and goat anti‐rabbit IgGs Fc‐HRP (1/10 000, AP156P; Millipore Corp.). Binding studies were carried by ELISA as described in Materials and methods. The absorbance of collagen‐bound VNs was corrected using the reactivity of each VN to anti‐VN IgGs. C, control VN incubated without enzyme (●); F, VN treated with fucosidase (▲); N, VN treated with neuraminidase (♦); F+N, VN treated with both fucosidase and neuraminidase (□); NG, VN treated with N‐glycosidaseF (○). (A) hVN, (B) pVN. Data are presented as mean ± SD.

Fig 5: Collagen‐binding activities of glycosidase‐treated VNs. Collagen type I was immobilized on wells of a 96‐well plate. The purified glycosidase‐treated VNs were added to the wells and reacted with the collagen in 10 mm Tris/HCl saline (pH 7.5). The VNs bound to collagen were detected by rabbit anti‐VN IgGs (1/25 000, LSL‐LB‐2096; Cosmo Bio Co., Ltd) and goat anti‐rabbit IgGs Fc‐HRP (1/10 000, AP156P; Millipore Corp.). Binding studies were carried by ELISA as described in Materials and methods. The absorbance of collagen‐bound VNs was corrected using the reactivity of each VN to anti‐VN IgGs. C, control VN incubated without enzyme (●); F, VN treated with fucosidase (▲); N, VN treated with neuraminidase (♦); F+N, VN treated with both fucosidase and neuraminidase (□); NG, VN treated with N‐glycosidaseF (○). (A) hVN, (B) pVN. Data are presented as mean ± SD.