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. Author manuscript; available in PMC: 2015 Apr 19.
Published in final edited form as: Biochem Pharmacol. 2014 Jul 24;91(3):369–379. doi: 10.1016/j.bcp.2014.07.013

Fig. 3.

Fig. 3

Binding of KCTD12 to the G-protein βγ subunits is modulated by receptor activation but not by PKA activation. (A) Scheme of the BRET Rluc-Gβ1 donor and Venus-KCTD12 acceptor pair. (B) BRET donor saturation curves from CHO-K1 cells stably expressing GB1 and GB2 and transiently expressing fixed amounts of Rluc-Gβ1 and Gγ2 together with increasing amounts of Venus-KCTD12. Following receptor activation with baclofen (100 µM, 5 min pre-incubation; grey traces) the BRET curves were shifted towards higher BRETmax values. Activation of PKA with 8-Br-cAMP (1 mM, 30 min pre-incubation; open circles) had no effect on the BRET curves. BRET is expressed as milli BRET units (mBU) determined as net BRET × 1000. Data points are means ± SD of technical quadruplicates of a representative experiment, n = 3.