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. Author manuscript; available in PMC: 2017 Nov 15.
Published in final edited form as: Biochemistry. 2015 Jul 24;54(30):4711–4730. doi: 10.1021/acs.biochem.5b00362

Figure 9. Recombinant CvGal2 binds to hemocyte extract.

Figure 9

(A) Hemocyte extract (1 μg/ml) coated wells were incubated with different concentration of rCvGal2 (0 to 30 μg/ml). Binding of rCvGal2 was measured in solid-based binding assay, and mean optical density at 450 nm (OD450nm) in triplicates with SEM was shown. (B) Hemocyte extract (1 μg/ml) coated wells were treated with neuraminidase (Ctrl) plus N-acetylgalactosidase (GalNac’ase), PNGase F (PNGF), O-glycosidase (OG’ase), or both PNGF and OG’ase. Binding of rCvGal2 to treated wells was measured as above. Relative binding to untreated cells was calculated, and the mean from triplicates with SEM is shown. *: p<0.05; ***: p<0.001 vs. control. (C) Hemocyte extract (1 μg/ml) (left panel) or PSM (50 ng/ml) (right panel) coated wells were pre-incubated with different concentration of anti-A or anti-B (1:25 to 1:1000 dilution) before incubation with rCvGal2 (2 μg/ml). Binding of rCvGal2 to treated wells was measured as above. Relative binding to untreated cells was calculated, and the mean from triplicates with SEM is shown. ***: p<0.001 vs. control.