Figure 1

Design and characterization of cell type specific Wnt signaling enhancing mimetic molecules. (A) Scheme of designed molecules. Fragments of RSPO2 spanning the Fu1 and Fu2 domains (action module) were fused the C-terminus of cell type specific or control scFv (targeting module). Specific mutations used are indicated. (B) In vitro STF activity of four RSPO proteins. Furin domains of human RSPO1-4 were tested as fusions to an anti-GFP scFv. RSPO2 furin ___domain alone (N37-E143) was used as a control. Luciferase activity unit is arbitrary (RLU). (C) STF activity of RSPO mimetics on HEK293 (top) or HuH-7 (bottom) in the presence (left) or absence (right) of an exogenous Wnt source (30% Wnt3a conditioned media). (D) Quantitative PCR analysis of ASGR1, ASGR2 and TFRC gene expression in HEK293, HuH-7 and A431 cells. The signals were relative to ACTB. (E) Western Blot analysis on LRP6 receptor and DVL2 phosphorylation in HuH-7 cells. Asterisk indicates a non-specific band that is detected in HuH-7 by the antibody used. The lower bands indicated by the arrow correspond to DVL2 with and without phosphorylation. Tubulin (TUB) is used as the loading control. 10% Wnt3a conditioned media was used as the exogenous Wnt source. Images were cropped for clarity. The uncropped images are presented in Fig. S2.