Fig. 1: Characterization of the monoFZD4-specific WNT/β-catenin signaling surrogate, L6-F4-2.
a Binding affinity of the recombinant F4-2_Fab molecule to FZD4 CRD measured by BLI assay. Dotted lines indicate the global fits generated by using a 1:1 Langmuir binding model. b Schematic of L6-F4-2 and the binding affinity of the L6-F4-2 molecule to the FZD4 CRD measured by BLI assay. Dotted lines indicate the global fits generated by using a 1:1 Langmuir binding model. c The binding specificity of L6-F4-2 against all ten FZD CRDs was examined by BLI assay. d Dose-dependent STF activity of L6-F4-2 or WNT3A in 293STF cells (top) or FZD4-transfected 293STF cells (bottom, OE = overexpression). e Dose-dependent STF activity of L6-F4-2 or Norrin in 293STF cells (top) or cells transfected with both FZD4 and LRP6 (bottom, OE = overexpression). f Dose-dependent STF activities of L6-F4-2 in HRMEC cells (top) and bEnd.3 cells (bottom). Inset box in the bEnd.3-STF graph is an enlarged plot of the Norrin response. g Quantitative RT-PCR of Axin2, Mki67, Lef1 and Mfsd2a gene expression in bEnd.3 cells. mRNA expression values were normalized by Actb gene expression. Results are from three independent experiments. Graphs are shown as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001, two- sided Mann–Whitney U test. Source data are provided as a Source Data file.
