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. 2012 Dec 4;288(5):3003–3015. doi: 10.1074/jbc.M112.418467

FIGURE 1.

FIGURE 1.

Regulation of AGS3-Rluc-Gαi1-YFP and AGS4-Rluc-Gαi1-YFP BRET by nonreceptor GEFs. AGS3-Rluc (10 ng) (A) and AGS4-Rluc (2 ng) (B) were expressed together with Gαi1-YFP (250 ng) in HEK cells, and BRET was measured as described under “Experimental Procedures.” Ric-8A, AGS1, or GIV(1660–1870) was expressed as indicated. Data are presented as the mean ± S.E. from four experiments with triplicate determinations. *, p < 0.05 compared with control. The relative fluorescent units and relative luciferase units for sample points in A and B are presented in Table 1. Lower panels, Ric-8A, AGS1, and GIV(1660–1870) immunoblots. Ric-8A and AGS1 proteins were detected with affinity-purified anti-Ric-8A and anti-AGS1 antibody, respectively. GIV(1660–1870) was detected with anti-His6 antibody. Each lane contains 50 μg of total protein, and the immunoblot is representative of three separate experiments. The GIV construct (pcDNA3.1/His::GIV(1660–1870)) encoded the carboxyl-terminal region of the protein as described under “Experimental Procedures.”