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. 2016 Aug 18;4(5):e00249. doi: 10.1002/prp2.249

Figure 7.

Figure 7

RGS14 accelerates the deactivation of Gαi1 proteins through its RGS domain following receptor activation. (A) HEK 293 cells were transfected with 200 ng of α2A‐adrenergic receptor, 200 ng of Venus‐Gβ1, 200 ng of Venus‐Gγ2, 25 ng of mas‐GRK3ct‐Luc, and 400 ng of Gαi1 and monitored for kinetic BRET as in Figure 1. Deactivation curves were normalized to maximum BRET and fit using a single exponential decay function in cells expressing Gαi1 and either 0 ng or 30 ng FLAG‐RGS14‐WT (n = 3). (B) Deactivation curves as in (A) in cells expressing Gαi1 and either 0 ng or 30 ng FLAG‐RGS14‐E92A/N93A (n = 3). (C) Deactivation curves as in (A) in cells expressing Gαi1 and either 0 ng or 30 ng FLAG‐RGS14‐Q515A/R516A (n = 3). (D) Deactivation rates (k deactivation) determined from curves fit in (A, B, and C). Error bars represent ± S.E. Statistical analysis was performed using a one‐way analysis of variance (ANOVA) with Dunnett's post hoc test (**P < 0.01, ***P < 0.001). RGS, Regulators of G protein signaling.