FIGURE 1.

Regulation of surface MC2 and MC5 receptors by MRAP. Surface expression of HA-MC2 (A), HA-MC5 (B), or HA-β2-adrenergic receptor (β2ADR) (C) measured by ELISA in non-permeabilized CHO cells using anti-HA antibody, as a function of the total expression of MRAP-V5 measured in parallel dishes by ELISA in permeabilized cells using anti-V5 antibodies. D, detection of MC2R, MC5R, MRAP, and MRAP2 mRNA in mouse adrenal glands or mouse Y1 adrenal cell line mRNA extracts by RT-PCR. Cells expressing HA-MC2 (E) or HA-MC5 (F) receptor in the presence or absence of MRAP-V5 were lysed, and receptors were immunoprecipitated with anti-HA antibody and resolved on SDS-PAGE. Blots were probed with anti-HA antibody. *, IgG heavy chain.