Figure 5

QRT-PCR for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E₁, E₂, and D₂. THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE₁ (0.1 μM) + NS398 (50 μM), (4) PGE₂ (0.1 μM) + NS398 (50 μM), (5) PGE₁ (0.1 μM) + PGE₂ (0.1 μM) + NS398 (50 μM), and (6) PGD₂ (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E₁, E₂, and D₂. THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE₁ (0.1 μM) + NS398 (50 μM), (4) PGE₂ (0.1 μM) + NS398 (50 μM), (5) PGE₁ (0.1 μM) + PGE₂ (0.1 μM) + NS398 (50 μM), and (6) PGD₂ (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. **p < 0.01 compared to NS398 (n = 5). ABCA1, ATP-binding cassette transporter A1; COX, cyclooxygenase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NS398, N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; PG, prostaglandin; QRT-PCR, quantitative real-time polymerase chain reaction.