Confocal analysis of cells was performed using a Leica SP8 microscope and 63× water objective. Coverslips with the cells were mounted using an Attofluor holder (Molecular Probes). To analyze the expression of the receptor constructs and DVL2 recruitment to the membrane, cells were maintained in imaging buffer. CFP was excited using a diode laser at 442-nm laser line, and fluorescence intensities were detected from 450 to 500 nm. GFP was excited using an argon laser at 488-nm laser line and detected at 509 to 600 nm. Images were acquired using 512 × 512 resolution, 400 Hz, a line average of 3, and a frame accumulation of 2. Confocal analysis of the fixed samples was done using a 63× oil-immersion objective. DAPI was excited using a diode 405 laser at 405-nm laser line, and fluorescence intensities were detected at 431 to 480 nm. Cy3 was excited using a diode-pumped solid-state laser at 561-nm laser line and detected at 590 to 679 nm. Images were acquired using 1024 × 1024 resolution, 400 Hz, and a line average of 8.

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