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. Author manuscript; available in PMC: 2022 Jun 4.
Published in final edited form as: Mol Cell. 2021 Sep 27;81(22):4605–4621.e11. doi: 10.1016/j.molcel.2021.09.007

Figure 3. D6R and C5aR2 exhibit a distinct GRK preference for βarr recruitment.

Figure 3

(A and B) HEK293 cells lacking indicated GRKs were transfected with the indicated receptor and βarrs followed by measurement of agonist-induced βarr recruitment by using the NanoBiT assay. Data (mean ± SEM) from three independent experiments are normalized with respect to basal signal (i.e., vehicle treatment).

(C) HEK293 cells expressing D6R were stimulated with CCL7 (100 nM), and total receptor phosphorylation was assessed by using the pIMAGO phospho-protein detection kit. A representative blot and densitometry-based quantification (mean ± SEM) from five independent experiments normalized with respect to basal signal are presented.

(D) Carboxyl-terminal truncation reduces the βarr2 interaction with D6R as assessed by a co-immunoprecipitation experiment using HEK293 cells expressing the indicated receptor construct and βarr2 followed by western blotting. A representative blot and densitometry-based quantification (mean ± SEM), normalized with D6RWT stimulation condition (treated as 100%), is presented (n = 3; one-way ANOVA; p < 0.001). See also Figures S3 and S4.