Figure 2
The effects of TMP on expression and activity of COX-2. RAW 264.7 cells were labeled overnight with [3H]-arachidonic acid, washed, then either left untreated (Media), or treated with LPS (1 μg/ml) and/or TMP (25 μM) for 16 h (A). Supernatants were collected and radioactivity determined by scintillation counting. RAW 264.7 cells were treated with LPS (1 μg/ml) alone or in combination with TMP (25 μM) and copy number of COX-2 mRNA determined by q-RT-PCR as described in the Materials and Methods (B). RAW 264.7 cells were treated with LPS (1 μg/ml) and/or TMP (25 μM), and the expression of COX-2 protein was examined by Western blot (C). TMP was added to purified placental ovine COX-2 protein and specific activity determined as described in the Materials and Methods (D).