Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025:2904:11-19.
doi: 10.1007/978-1-0716-4414-0_2.

Visualization and Quantification of NFAT1 Translocation in T Cells

Franziska Möckl  1 , Dejan Kovacevic  1   2   3   4 , René Werner  2   3   4 , Björn-Philip Diercks  5 , Mariella Weiß  6
Affiliations

Visualization and Quantification of NFAT1 Translocation in T Cells

Franziska Möckl et al. Methods Mol Biol. 2025.

Abstract

NFAT1 translocation to the nucleus and consequent IL-2 production in T cells can be used as a downstream read-out for initial T cell activation via the T cell receptor complex. Here, we describe a protocol for the detection of NFAT1 translocation using anti-CD3/anti-CD28 coated beads as a stimulus of primary murine CD4+ T cells by immunohistochemistry. Further, an open-source Python code is described to determine NFAT1 translocation.

Keywords: Calcium; Immunofluorescence; Nuclear Factor of Activated T cells (NFAT); Nuclear translocation; Python; T cells.

PubMed Disclaimer

Similar articles

See all similar articles

References

    1. Macian F (2005) NFAT proteins: key regulators of T-cell development and function. Nat Rev Immunol 5:472–484. https://doi.org/10.1038/nri1632 - DOI - PubMed
    1. Lee J-U, Kim L-K, Choi J-M (2018) Revisiting the concept of targeting NFAT to control T cell immunity and autoimmune diseases. Front Immunol 9:2747 - DOI - PubMed - PMC
    1. Luo C, Burgeon E, Carew JA et al (1996) Recombinant NFAT1 (NFATp) is regulated by calcineurin in T cells and mediates transcription of several cytokine genes. Mol Cell Biol 16:3955–3966. https://doi.org/10.1128/MCB.16.7.3955 - DOI - PubMed - PMC
    1. Kar P, Lin Y-P, Bhardwaj R et al (2021) The N terminus of Orai1 couples to the AKAP79 signaling complex to drive NFAT1 activation by local Ca2+ entry. Proc Natl Acad Sci 118:e2012908118. https://doi.org/10.1073/pnas.2012908118 - DOI - PubMed - PMC
    1. Son G-Y, Subedi KP, Ong HL et al (2020) STIM2 targets Orai1/STIM1 to the AKAP79 signaling complex and confers coupling of Ca2+ entry with NFAT1 activation. Proc Natl Acad Sci 117:16638–16648. https://doi.org/10.1073/pnas.1915386117 - DOI - PubMed - PMC

Substances

LinkOut - more resources