Intracellular Ca2+ mobilization was measured using the FLIPR Calcium 5 Assay Kit (Molecular Devices). Briefly, HPAF-II cells (350,000 cells/ml) were seeded (48 hours before the assay) onto poly-d-lysine–coated black 96-well culture plates with glass bottom. Cells were loaded with FLIPR calcium 5 indicator dye (Molecular Devices) and incubated with DMSO or Gαq inhibitor (YM-254890; 100 nM) for 60 min and then stimulated with recombinant WNT-5A (2000 ng/ml). Intracellular Ca2+ mobilization was detected with the FlexStation II Benchtop Multi-mode Microplate Reader (Molecular Devices).

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