Figure 5.

Preparation and Wnt3A antagonistic function assay of Sizzled and its derivatives. A, size-exclusion chromatography profiles of Sizzled (blue, 1), Sizzled C115S/C156S (pink, 2), Sizzled C115S/C156S/H116Y/H118F (red, 3), Sizzled CRD C115S (cyan, 4), and Sizzled CRD C115S/H116Y/H118F (black, 5) alongside an inset showing SDS-PAGE analysis of peak fractions. B, sequence alignment of thumb and index finger motifs of XWnt8, XWnt3a (X. laevis Wnt3A), and hWnt3a (human Wnt3A). C, fold changes of TOPflash/FOPflash firefly luciferase activity (each value normalized by Renilla luciferase activity) under different experimental conditions. For each protein, 3 columns from left to right, respectively, correspond to protein concentrations of 2.56, 25.6, and 256 nm (represented by L, M, and H, respectively). ^∧∧, p value < 0.05, determined through comparing results in the presence of Wnt3A with the vehicle control; *, p value <0.05, and **, p value < 0.01, comparing results in the presence of the indicated protein sample with those obtained in the presence of Wnt3A alone.