MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT EP3 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human EP3 cDNA (GenBank Accession Number NM_000957) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM-F12, 10% FBS, 10 µg/mL puromycin
Stable after minimum two months continuous growth
Background: Prostaglandin E2 (PGE2) is involved in a number of physiologic and pathophysiologic events in many tissues of the body. The biologic effects of PGE2 are mediated through interaction with specific membrane-bound G protein-coupled prostanoid EP receptors. EP3 receptor (or PTGER3) is expressed as multiple transcripts through alternative splicing, with each transcript showing a different tissue-specific distribution. PGE2 may mediate fever generation in response to both endogenous and exogenous pyrogens by acting at the EP3 receptor. EP3-mediated neuronal pathways converge at corticotropin-releasing hormone containing neurons in the paraventricular nucleus of the hypothalamus to induce HPA axis activation during sickness.
At cellular level, EP3 has been shown to couple to both Gs and Gi/o types of the heterotrimeric G proteins to stimulate or inhibit intracellular cAMP synthesis. This cell line does not respond to pertussis toxin (PTX) treatment, indicating its coupling to Gi/o proteins is insignificant (See Figure).
Application: Functional assays
Figure 1. Dose-dependent stimulation of intracellular cAMP accumulation upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). Figure 2. Receptor expression on cell surface measured by flow cytometry (FACS) using an anti-FLAG antibody. Thin line: parental cells; thick line: receptor-expressing cells.
Adam et al. (1994) Cloning and expression of three isoforms of the human EP(3) prostanoid receptor. FEBS Lett 338:170-174.
Matsuoka et al. (2003) Impaired adrenocorticotropic hormone response to bacterial endotoxin in mice deficient in prostaglandin E receptor EP1 and EP3 subtypes. Proc Nat Acad Sci USA 100:4132-4137.
Ushikubi et al. (1998) Impaired febrile response in mice lacking the prostaglandin E receptor subtype EP(3). Nature 395:281-284.