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Stable HeLa Luciferase Reporter Cell Lines Expressing GAL4 Fusion,,,Transactivators

s five times higher than the control, when cells were treated with 100 ng/ml of EGF, and almost 50-fold higher when cells were challenged with PMA (60 ng/ml). Dulbeccos modified Eagles medium (DMEM) and dimethylsulfoxide (DMSO) were used as negative controls.

Fig.4

For the final testing, we confirmed purity by screening for mycoplasma and established viability after freezing in liquid nitrogen. The HLR, HLR-CREB, HLR-Elk1, and HLR-cJun cell lines are available as frozen stocks containing 5 x 105 cells; each cell line includes the corresponding positive and negative control plasmids.

Conclusions

The HLR-CREB, HLR-Elk1, and HLR-cJun stable cell lines offer a new approach and added convenience for studying the intricate interactions of signal transduction pathways. These cell lines can be used to evaluate whether a new gene product or extracellular stimulus is involved in signaling pathways that converge at CREB, Elk1, and c-Jun transcription factors; furthermore, the cell lines can be used to identify which step along the pathway is targeted. The HLR-CREB, HLR-Elk1, and HLR-cJun cell lines can be adapted for high-throughput screening of inhibitors or activators of these signaling pathways. In addition, the HLR cell line can be used in a variety of other applications, such as assessing protein-protein interactions in GAL4-derived mammalian two-hybrid systems.

REFERENCES
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  3. Treisman, R. (1996) Curr. Opin. Cell Biol. 8: 205-215.

  4. Xu, L., et al. (1997) Strategies 10: 1-3.

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