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. Author manuscript; available in PMC: 2010 Nov 28.
Published in final edited form as: Sci Signal. 2010 Apr 6;3(116):ra28. doi: 10.1126/scisignal.2000593

Fig. 1.

Fig. 1

Effects of MRAP and MRAP2 on MC2R and MC4R signaling. (A to E) CHO cells were transfected with plasmids encoding HA-hMC2R or HA-hMC4R and V5-mMRAP-FLAG, V5-mMRAP2-FLAG, or V5-hMRAP2-FLAG and (C and D) CRE-Luc. (A) cAMP accumulation in cells transfected with plasmid encoding HA-hMC2R that were incubated for 20 min with 0.1 mM isobutylmethylxanthine and either vehicle or ACTH (1–39). CRE-Luc activity in cells transfected with plasmid encoding HA-hMC2R (C) or HA-hMC4R (D) that were incubated for 4 hours with vehicle, ACTH(1-39), NDP-α-MSH, or forskolin (20 μM). Data are normalized to the forskolin response. ACTH(1-39) and NDP-α-MSH did not generate any responses in mock-transfected cells. The forskolin response in cells that had mMRAP2 averaged 69 ± 29% higher than that in cells that had mMRAP (n = 11 experiments). (B and E) Surface HA-hMC2Rs or HA-hMC4Rs in nonpermeabilized cells were detected by ELISA with an antibody against the HA tag. In (B), data are normalized to the value with MRAP, and in (E), data are normalized to the value with MC4R alone. Shown are the mean and SE or range from representative experiments performed in triplicate, for (A) through (D), or in duplicate, for (E). *P < 0.05 versus receptor alone.