Fig. 4: ZNRF3 dimerization through complexation with LGR4 and RSPO2 is important for Wnt/β-catenin signal potentiation.
From: Structural insights into Wnt/β-catenin signaling regulation by LGR4, R-spondin, and ZNRF3
a Pull-down assay of ZNRF3 (WT) or ZNRF3 (E95N/E97T) in the presence or absence of LGR4 and RSPO2. Expi293F cells were transfected with ZNRF3 ECD-TMD (Flag-tagged), ZNRF3 ECD-TMD (HA-tagged), LGR4 ECD-TMD (Myc-tagged), and RSPO2 Fu1-2 (Strep-tagged). Cell lysates were incubated with anti-FLAG affinity beads, and bound proteins were detected by western blotting. Data are representative of three independent experiments. b TOPflash reporter assays using full-length wild-type (WT) or dimerization-deficient mutant (E95N/E97T) ZNRF3. HEK293T cells were transfected with the indicated siRNA, followed by plasmid transfection, and then treated with or without 5% Wnt3a-conditioned medium in the presence or absence of 200 ng/mL RSPO2. EV: empty vector. n = 3 biological replicates. Bars represent mean ± standard error of the mean (SEM), and dots show individual data points. Statistical significance was determined using Two-way ANOVA with two-sided Tukey’s test. ***P < 0.001. ns not significant. Source data are provided as a Source data file.
