Extended Data Fig. 1: NTSR1–GRK2–Gαq complex assembly.
From: GPCR activation and GRK2 assembly by a biased intracellular agonist
a, Screening for GPCR–GRK2 complexes by tango assay. RLU, relative luciferase units, which was normalized to the values of NTSR1. Data were processed as mean ± S.D. from three independent experiments (n = 3), performed in triplicates. Statistical significance of differences between NTSR1 and other receptor was determined by two-sided one-way ANOVA. **P < 0.01 and *** P < 0.001 versus NTSR1 (P < 0.0001, <0.0001, <0.0001, <0.0001, <0.0001, <0.0001, <0.0001, <0.0001, <0.0001 from left to right). b, NTS and SBI-553 improve NTSR1-GRK2 interaction determined by Tango assay. Data were processed as mean ± S.D. from three independent experiments (n = 3), performed in triplicates. Statistical significance was determined by two-sided one-way ANOVA. **P < 0.01 and ***P < 0.001 (P = 0.0024, 0.0027, <0.0001, 0.0001, <0.0001, <0.0001 from left to right). c-e, SDS-PAGE of the complexes. NTSR1–GRK2–Gαq–Gβγ complex (c), NTSR1_LgBiT–GRK2_HiBiT–Gαq–Gβγ complex (d), NTSR1–GRK2–Gαq complex before crosslinking (left panel of e) and NTSR1–GRK2–Gαq complex crosslinked by BS3 (right panel of e). For gel source data, see Supplementary Data Fig. 1. Representative Figures from at least three independent experiments were shown. f, Size-exclusion chromatography elution profile of the NTSR1–GRK2–Gαq complex. Red star indicates the monomer peak of the complex. g, Cryo-EM micrograph of the NTSR1–GRK2–Gαq complex. Representative Cryo-EM micrograph from 57,477 movies was shown. Particles picked for 3D classifications were highlighted in red circles.
