Protocol for intranuclear Nrf2 detection in activated mouse, human, and Jurkat T cells by spectral flow cytometry

Nuclear factor erythroid 2-related factor 2 (Nrf2) coordinates cellular antioxidation responses. Its expression increases in T cells upon activation. Here, we present a protocol for quantitative detection of intranuclear Nrf2 in activated mouse, human, and Jurkat T cells using spectral flow cytometry. We describe procedures for primary T cell isolation, in vitro activation, intranuclear staining, and flow cytometric acquisition and analysis, including gating strategies for CD4⁺ and CD8⁺ T cells. For complete details on the use and execution of this protocol, please refer to Tripathi et al.¹.