Skip to main content
Figure 8 | BMC Developmental Biology

Figure 8

From: Structure and epitope distribution of heparan sulfate is disrupted in experimental lung hypoplasia: a glycobiological epigenetic cause for malformation?

Figure 8

Functional analysis of antibody epitope structures via competition ELISA with FGF2 and FGF9. FGF2 and FGF9 competed with a number of antibodies for HS binding, indicating that epitope structures are analogous to structures recognised by these growth factors. FGF2 competed for all six epitopes to variable extents, but most significantly with EV3C3V. FGF9, in contrast, showed more competitive selectivity and was only able to compete for two epitope structures, recognised by HS3B7V and HS3A8V. PMHS was biotinylated and immobilised on streptavidin coated microtitre plates. Equilibrium binding of HS antibodies in the presence of various concentrations of FGF2 (A, B) or FGF9 (C, D) were quantified at A490 using an anti-VSV-G tag antibody (P5D4) followed by HRP conjugated anti-mouse antibody and OPD substrate. Absorbance values were converted to % inhibition of antibody binding to HS by FGFs and plotted against FGF concentration (A, C). Curves were generated using OriginPro and a non-linear logistic dose response fit. IC50 values (B, D) were calculated as the concentration of FGF required for 50% inhibition of antibody binding to immobilised HS. Values are the mean of triplicate samples, error bars represent the S.E and the data are representative of two separate experiments. * denotes antibody epitopes not competed for by FGF.

Back to article page