Figure 3: mGpr83 forms heterodimers with mGhsr and modifies its signalling properties. | Nature Communications

Figure 3: mGpr83 forms heterodimers with mGhsr and modifies its signalling properties.

From: The orphan receptor Gpr83 regulates systemic energy metabolism via ghrelin-dependent and ghrelin-independent mechanisms

Figure 3

Cells were transiently cotransfected with equal amounts of differentially tagged Gpr83 and Ghsr, respectively. Sandwich ELISA studies in COS-7 cells, expressed as % absorption (492 nm/620 nm) relative to positive control (NHA-Mc3r+Ghsr-Flag) and mock (NHA-Ghsr) (a). YFP-based protein complementation assay in HEK293 cells, expressed as % fluorescent cells to positive control (Mc3r-YFP1+Ghsr-YFP2) (b) and functional characterization of the Ghsr/Gpr83 heterodimer in HEK293 (c). Measurement of inositol-3 phosphate (IP3) following stimulation with 1,000 nM ghrelin was carried out 48 h after transfection. (d) HA-tags of receptors on the cell surface in comparison with untagged rM3R as negative control. (e) HA- and Flag-double tagged receptors expressed as fold over the rM3R negative control (e,d). Data were assessed from a minimum of three independent experiments (a–e), each performed at least in triplicates (a,c–e). Cntrl, control; rM3R, rat muscarinic acetylcholine receptor M3. Data represent mean±s.e.m. ***P<0.001 based on one-way analysis of variance followed by Bonferroni’s post-hoc test.

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