Figure 3.

The GPR124 C1196Y mutation reduces Wnt signaling activity. A, Schematic illustration of the C1196Y alteration in the intracellular domain (ICD) of GPR124. B, Wild‐type (WT) GPR124 and GPR124 C1196Y were cloned into pcDNA3.1‐3xFLAG construct and overexpressed in HEK 293T cells. Protein subcellular location was determined by confocal immunofluorescence staining. C, Expression levels of GPR124 WT, GPR124 C1196Y, and GPR124 with ICD deletion (GPR124 △ICD) were determined by Western blotting. D‐E, Wnt signaling activity was measured by AXIN2 qRT‐PCR (D) and β‐catenin‐dependent transcription assay TOP‐Flash (E) after transfecting HEK 293T cells with the indicated plasmids. Data are presented as mean ± SE. *P < .05, **P < .01. NS, not significant, Student's t test