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, j BRET2 assay to evaluate PrRP20-induced dissociation of heterotrimeric G q protein ( i ) and G i protein ( j ) (Δspan = the span value of PrRP20 to a specific PrRPR variant – the span value of PrRP20 to WT PrRPR; yellow column means Δspan ≤ –50%; blue column means Δspan ≤ –80%). Data are presented as means ± SEM; n = 3 independent samples, each consisting of triplicate measurements; significance was determined by one-way ANOVA with Dunnett’s multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001. Exact span values are provided in Supplementary Table S3 .

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