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Prostaglandin synthases are key enzymes in the prostanoid biosynthetic pathway. Arachidonic acid released from membrane phospholipid stores by the activity of phospholipases is converted to PGH2, a common precursor of prostaglandins, by the enzymatic activity of prostaglandin synthase (PGS). There are two forms of PGS enzymes; PGS1 and PGS2. PGS1 is constitutively expressed in almost all cell types, whereas PGS2 is induced in a number of cell types by appropriate stimuli. This chapter describes methods used for the detection of PGS2 expression by western and northern analyses in murine Swiss 3T3 fibroblasts. These methods have also been used successfully to detect PGS2 expression in a number of other murine cell lines (Table 1 ).
Table 1Induction of Murine Cell Lines
Murine cells in which PGS2 is induced | Inducer | Reference |
|---|---|---|
RAW 264.7 (macrophages) | LPS | 1 |
MMC-34 (mast cells) | IgE-anti IgE | 2 |
Bone marrow derived mast cells | IgE-anti IgE | 3 |
Peritoneal macrophages | LPS, Adrenelectomy | 4 |
Primary embryonic neuronal cultures | BDNF, NT-3 | 5 |
Murine embryonic fibroblasts | TPA, TGF-beta | 6 |
3T3 Fibroblasts | TPA, Serum | 7 |
