Cellular Ca2+-Responding Nanoluciferase Reporter Gene System Directed by Tandemly Repeated Pseudo-palindromic NFAT-Response Elements
Luciferase reporter gene systems based on the NFAT-response element (RE) have been used to monitor intracellular Ca2+ elevation. However, Ca2+ mobilization agent (e.g., ionomycin) alone is not adequate to activate the currently often employed reporter gene that contains the NFAT-RE found in the IL2 promoter. In addition to activation of NFAT through the Ca2+-calmodulin/calcineurin pathway, activation of AP-1 as a partner transcription factor is essential for the IL2-based NFAT-RE system. Here, we describe a detailed method for the recently developed new reporter gene system containing the NFAT-RE from the IL8 promoter. This system enables us to monitor endpoint effects of Ca2+-mobilizing agonists independent of AP-1 activation.
Key wordsCalcineurin Calcium signaling Calmodulin Carbachol Luminescence NanoLuc NFAT Reporter gene assay Transcription factor
This work was supported by JSPS KAKENHI Grant Number 17H03803 (Grant-in-Aid for Scientific Research (B) to M.M.). We thank current and past members of the Laboratory of Molecular and Cellular Regulation for valuable suggestions and discussion.
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