Calcium/Calcineurin, T Cell Activation

Nuclear Factor of Activated T cells (NFAT) are a family of transcriptional factors that play an important role in immune response as well as in the development of cardiac, skeletal muscle, and nervous systems.  NFATs are regulated by calcium signaling. Through calcium activation of the phosphatase calcineurin, NFATc proteins translocate from the cytoplasm into the nucleus, where they cooperate with other proteins to mediate gene expression. Nuclear import of NFAT is blocked by kinases such as PKA and GSK3. NFATs are also implicated in breast cancer. 

 

Signosis has established NFAT luciferase reporter cell lines that has been stably transfected with a NFAT-luciferase reporter construct. Via the analysis of luciferase, the cell line can be employed to monitor the cellular changes of NFAT activities that are triggered by stimulation, compound treatment, enforced gene expression or gene knockdown.  

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The cell line is established by transfection using a pTA-NFAT-luciferase reporter vector, which contains NFAT binding sites, a minimal promoter upstream of the firefly luciferase coding region, along with hygromycin expression vector followed by hygromycin selection.  The hygromycin resistant clones were subsequently screened for PMA+ionomycin-induced luciferase activity.