Figure 4.
Dkk4 binds with high affinity to LRP6 E1E2. A, schematic representations of the human His-tagged Dkk4FL and Dkk4N and LRP6 E1E2-Fc expression constructs. B, Western blot analysis of pulldown assays, illustrating the binding of His-tagged Dkk proteins to LRP6 E1E2-Fc captured on protein A beads. C, representative biolayer interferometry sensorgrams for His-tagged Dkk4FL binding to immobilized LRP6 E1E2-Fc. Association and dissociation phases of sensorgrams are shown for a range of indicated concentrations of His-tagged Dkk4FL, together with the steady-state binding curve derived from the maximum response (Rmax) observed. The KD reported was obtained by fitting to a one-site binding model using Prism version 6.0. D, a summary of biolayer interferometry KD measurements from two individual experiments and representative of a minimum of three or more independent measurements. Errors shown are the S.E. calculated for individual fitted curves using Prism version 6.0. ND, no data obtained.