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. 2018 Jul 17;11:245. doi: 10.3389/fnmol.2018.00245

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Copyright © 2018 Xue, Bai, Ji, Chen, Wang, Wang, Yang, Zhang, Jiang, Pan, Cheng and Chen.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Förster resonance energy transfer assay to detect the dimerization of GHSR1a/OX1R. (A) FRET imaging of constitutive GHSR1a/OX1R heteromeric interactions in living cells. HEK293 cells were transiently transfected with plasmids encoding (a) as a negative control group, CRH1R-ECFP and GHSR1a-EYFP. (b) GHSR1a-EYFP (acceptor), (c) OX1R-ECFP (donor), (d) OX1R-ECFP and GHSR1a-EYFP. Left panels, ECFP; center panels, EYFP; right panels, corrected FRET. (B) Normalized FRET values, calculated as described in Experimental Procedures. The data represent mean ± SEM (N = 3) of four independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison post hoc test. ^∗∗∗p < 0.001 vs. the negative groups CRH1R-ECFP and GHSR1a-EYFP.