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. 2017 Feb 1;37(5):1162–1175. doi: 10.1523/JNEUROSCI.2181-16.2016

Figure 5.

Figure 5.

KCTD homo- and hetero-oligomers bind to both GABAB receptors and G-proteins in live HEK293 cells. A, BRET between KCTD oligomers and GABAB receptors in living cells. Cells were transfected with fixed amounts of NTRlucKCTD and CTRlucKCTD constructs and increasing amounts of Myc-GABAB1 and HA-GABAB2-GFP constructs. BRET was measured between reconstituted Rluc and GABAB2-GFP. Data points represent the mean of technical duplicates combined from 6 or 7 independent experiments. Donor/acceptor ratios needed to reach half BRET saturation (BRET50) were as follows: KCTD12 homo-oligomer: 0.27 ± 0.03; KCTD16 homo-oligomer: 2.55 ± 0.97; KCTD12/KCTD16 hetero-oligomer: 1.91 ± 0.29; KCTD16/KCTD12 hetero-oligomer: 6.65 ± 0.71 (mean ± SEM, n = 6 or 7). B, BRET between KCTD oligomers and the G-protein in live cells. Cells were transfected with fixed amounts of NTRlucKCTD and CTRlucKCTD constructs and increasing amounts of Gβ2 and Gγ2-YFP constructs. BRET was measured between reconstituted Rluc and Gγ2-YFP. Data points represent the mean of technical quadruplicates from a representative experiment (n = 3).