CHO cells were seeded in six-well plates and transfected with the indicated plasmids. Cells were then lifted using TrypLE Express (Thermo Fisher Scientific) and resuspended in 500 μl of DMEM/F12. Cell suspension (7 μl/well) was transferred in a white opaque 384-well plate. Agonist solution (7 μl) diluted in IP-One kit stimulation buffer containing lithium was then added to each well and incubated for 1 hour at 37°C. Cells were then lysed, and accumulated IP1 was measured using the IP-One kit (Cisbio) following the manufacturer’s instruction. Readings were performed on a SpectraMax i3 plate reader (Molecular Devices). Each condition was run in triplicate, and experiments were repeated independently at least three times.

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