Table 1.
Amplification of LPS-stimulated gene expression by PA
| CT in Control Cells | CT in LPS-Treated Cells | CT in PA-Treated Cells | CT in LPS and PA-Treated Cells | Ratio of LPS-Treated vs. Control | Ratio of PA-Treated vs. Control | Ratio of LPS and PA-Treated vs. Control | |
|---|---|---|---|---|---|---|---|
| Cytokines | |||||||
| IL-6 | 30.55 | 30.39 | 29.54 | 25.86 | 1.1 | 2.0 | 25.8 |
| TNFα | 23.96 | 22.32 | 23.33 | 20.43 | 3.1 | 1.5 | 11.6 |
| IL-1α | 30.88 | 29.32 | 30.05 | 24.33 | 2.9 | 1.8 | 94.0 |
| IL-1β | 30.33 | 27.94 | 30.09 | 24.75 | 5.2 | 1.2 | 47.7 |
| IL-2 | 31.3 | 31.15 | 29.77 | 27.56 | 1.1 | 2.9 | 13.4 |
| CSF2 | 32.53 | 30.31 | 30.11 | 27.91 | 4.7 | 5.4 | 24.6 |
| CSF3 | 31.04 | 28.83 | 29.30 | 21.64 | 4.6 | 3.3 | 676.9 |
| TNFβ | 30.52 | 29.87 | 29.71 | 27.47 | 1.6 | 1.8 | 8.3 |
| IFNβ | 31.99 | 31.83 | 30.39 | 28.49 | 1.1 | 3.0 | 11.3 |
| IFNγ | 31.63 | 30.83 | 30.52 | 27.93 | 1.7 | 2.2 | 13.0 |
| MCP-1 | 28.41 | 24.71 | 27.82 | 20.27 | 13.0 | 1.5 | 281.9 |
| IL-10 | 30.22 | 29.45 | 29.30 | 27.25 | 1.7 | 1.9 | 7.8 |
| IL-12α | 31.26 | 30.73 | 30.58 | 28.33 | 1.4 | 1.6 | 7.6 |
| COX-2 | 28.1 | 25.55 | 27.25 | 20.95 | 5.8 | 1.8 | 141.5 |
| TLR and TLR-related signaling molecules | |||||||
| TLR3 | 27.58 | 26.95 | 27.14 | 26.16 | 1.5 | 1.4 | 2.7 |
| TLR5 | 31.92 | 30.99 | 29.94 | 27.62 | 1.9 | 3.9 | 19.6 |
| MD-2 | 25.16 | 25.13 | 25.24 | 24.05 | 1.0 | 0.9 | 2.2 |
| Irak2 | 24.82 | 24.18 | 24.92 | 23.50 | 1.6 | 0.9 | 2.5 |
| TRIF | 25.72 | 25.21 | 25.34 | 24.02 | 1.4 | 1.3 | 3.2 |
| IkBβ | 29.04 | 29.41 | 28.55 | 26.52 | 0.8 | 1.4 | 5.7 |
| CD80 | 27.79 | 26.43 | 27.49 | 24.72 | 2.6 | 1.2 | 8.4 |
| CD86 | 31.1 | 29.97 | 29.93 | 25.90 | 2.2 | 2.3 | 36.7 |
| Transcription factors | |||||||
| Jun | 24.89 | 24.77 | 24.73 | 23.28 | 1.1 | 1.1 | 3.1 |
| C/EBP | 22.45 | 21.71 | 22.10 | 20.76 | 1.7 | 1.3 | 3.2 |
| PPARα | 30.91 | 30.42 | 29.51 | 27.59 | 1.4 | 2.6 | 10.0 |
| Housekeeping gene | |||||||
| GAPDH | 17.86 | 17.74 | 17.81 | 17.88 | 1.1 | 1.0 | 1.0 |
PA, palmitic acid; CT, threshold cycle; CSF, colony-stimulating factor; MCP-1, monocyte chemotactic protein-1; COX-2, cyclooxygenase-2; TLR, Toll-like receptor; MD-2, myeloid differentiation factor-2; Irak2, IL-1 receptor-associated kinase 2; TRIF, TIR-domain-containing adapter-inducing IFNβ; C/EBP, CCAAT enhancer-binding protein; PPARα, peroxisome proliferator-activated receptor-α. RAW 264.7 cells were treated with 1 ng/ml LPS, 100 μM PA, or both for 24 h. After treatment, cells were harvested and subjected to PCR array study, as described in materials and methods. The GAPDH mRNA was used as a housekeeping gene. To compare the gene expression in cells treated with LPS, PA, or LPS + PA with that in control cells, ΔCT was first calculated by the following formula: ΔCT = CT in LPS, PA, or LPS + PA-treated cells − CT in control cells. The gene expression in cells treated with LPS, PA, or LPS + PA was presented as the fold of the control gene expression and calculated as 2ΔCT.