Table 1. The reporter gene and positive control plasmids in reporter gene assay.
| Singnaling Pathway | Wnt | C/EBP | GAS | NF-κB | AP-1 |
|---|---|---|---|---|---|
| Reporter gene plasmid | pTop-flash | pC/EBP | pGAS | pNF-κB | pAP-1 |
| Positive control plasmid | pβ-catenin | pPKA | pIFN-γ | pTNF-α | pMEKK |
| Relative luciferase unit | 12.7691 | 0.232175 | 0.172731 | 0.272368 | 1.295776 |
| Positive control fluorescence intensity | 25.69433 | 2.103423 | 3.249269 | 1.957258 | 158.182 |
| Negative control fluorescence intensity | 3.478703 | 0.095596 | 0.101221 | 0.177325 | 1.032595 |
| Relative activation times | 3.67065 | 2.42871 | 1.70647 | 1.53598 | 1.25487 |
Cells were harvested at 48 h after transfection, lysed and analyzed using the Dual-Luciferase Reporter Assay System according to the manufacturer's protocol. Firefly luciferase intensity divided by renilla luciferase intensity as a relative luciferase unit (RLU). The relative activation times was the ratio that calculated between RLU and negative control. The signaling pathway was activated if activation multiple more than three times. All reporter experiments were performed in triplicate and repeated three times and the data were expressed as the mean ± SD.