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Cells were transiently transfected using polyethyleneimine (PEI) with 0.25 g of plasmid DNA per well (WT human GIP receptor (E354 variant), human GIP receptor E354Q variant, or empty vector (pcDNA3.1+)) as previously described ( 2.4 cAMP measurement cAMP accumulation in Cos7 cells transfected with the WT or E354Q GIP receptors was measured using the LANCE cAMP detection kit (PerkinElmer Life and Analytical Sciences, MA, United States) as described previously ( 2.5 IP 1 measurement The IP-one Gq assay kit (Cisbio, PerkinElmer) was used to quantify accumulated myo-inositol-1-phosphate (IP 1 ), a by-product of IP3 produced after receptor-mediated G q activation in Cos7 cells, as previously described ( 1
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