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. 2015 Jul 30;5:12584. doi: 10.1038/srep12584

Figure 4. HEK293 cells were transiently transfected with 0.2 μg GPR103-Rluc or 0.2 μg OX1R -Rluc and 0.6 μg OX1R-EGFP or 0.6 μg GPR103-EGFP.

Figure 4

(a) ***p < 0.001 to control OX1R-Rluc + EGFP (n = 3), ###p < 0.001 to control GPR103-Rluc+EGFP (n = 3). HEK293 cells were co-transfected a constant amount of cDNA for GPR103-Rluc or OX1R-Rluc (0.15 μg) and increasing concentration of EGFP-tagged cDNAs. With the increasing expression (0.15–0.75 μg) of OX1R-EGFP or GPR103-EGFP, the saturation curve indicated the specific interaction between GPR103 and OX1R (b) GPR103-Rluc and OX1R-EGFP (1:3 and vice versa) were co-transfected into HEK293 cells and mBRET was measured upon treatment with OXA or QRFP (10 nM; n = 3). *p < 0.05, **p < 0.01 compared to basal unstimulated levels (c) HEK293 cells were transiently transfected with GPR103-Rluc, OX2R-EGFP or Rluc (all at 0.2 μg) and 0.6 μg OX2R-EGFP, or 0.6 μg EGFP (n = 3). ***p < 0.001 to controls GPR103-Rluc+EGFP and OX2R-EGFP+Rluc (d) HEK293 cells were co-transfected a constant amount of cDNA for GPR103-Rluc (0.15 μg) and increasing concentration (0.15–0.75 μg) of EGFP-tagged cDNAs. With the increasing expression of OX2R-EGFP and GPR103-Rluc, the saturation curve indicated a specific interaction between GPR103 and OX2R (e) GPR103-Rluc and OX2R-EGFP (1:3) were co-transfected into HEK293 cells and treated with OXA, OXB and QRFP (10 nM; n = 3). *p < 0.05, **p < 0.01 compared to basal levels (f).