HPAF-II and PANC-1 cells were seeded at 1000 to 2000 cells per well in 96-well plates. The next day, cells were treated with the PORCN inhibitor C59, the Gαq inhibitor FR900359, or vehicle in technical triplicate at the indicated concentrations. Medium containing inhibitor or vehicle was replaced every 3 days. Cell viability was measured using Alamar Blue (Thermo Fisher Scientific, catalog no. DAL1025) approximately 6 to 7 days after seeding following the manufacturer’s instructions. Fluorescence was measured at 560-nm excitation and 590-nm emission with a SPECTRAmax GEMINI Microplate Spectrofluorometer plate reader (Molecular Devices).

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