Table 1. Comparison of values for HA-hMC4R and hMC4R coupling to adenylyl cyclase in HEK293 cells shown in Fig 1 and S1 Fig For HA-hMC4R + pcDNA 3.1, hMRAPα-FLAG or hMRAP2-FLAG, normalised adenylyl cyclase data are shown as mean ± s.e.m for baseline and maximum responses and EC₅₀ values are shown with 95% confidence intervals in brackets.

For hMC4R + pcDNA 3.1 or hMRAP2, normalised adenylyl cyclase data are shown as mean ± s.e.m for baseline values only. Statistically significant differences for HA-hMC4R or HA-hMC4R + pcDNA 3.1 were * p < 0.05; *** p < 0.009; **** p < 0.0001. For direct comparison of hMC4R-eGFP and hMC4R coupling to adenylyl cyclase, mean values for raw data are shown ± s.e.m for baseline and maximum responses and EC₅₀ values are shown with 95% confidence intervals in brackets.

Transfection	Baseline	Maximum	EC50 (nM)
HA-hMC4R + pcDNA 3.1	0.224 ± 3.064	100.4 ± 2.008	1.560 (1.186, 2.052)
HA-hMC4R + hMRAPα-FLAG	17.65 ± 7.065*	116.6 ± 3.062**	0.388 (0.235, 0.641)***
HA-hMC4R + hMRAP2-FLAG	-1.182 ± 9.402	148.9 ± 4.527***	0.619 (0.388, 0.988)**
hMC4R + pcDNA 3.1	-0.473 ± 4.720
hMC4R + hMRAP2	0.321 ± 5.008