Flow Cytometry Analysis of IFN-γ in Jurkat T Cells

The cells were dispersed into a single-cell suspension and resuspended in staining buffer (BD, New Jersey, USA) while T cells (Jurkat) were treated with PE-CD3 (130-117-139, 1 : 50, Miltenyi Biotec, Bergisch Gladbach, Germany), fixed, permeabilized, and stained with Pacific blue-IFN-γ (BioLegend, San Diego, CA, #505817, rat, 1 : 50), followed by detection utilizing flow cytometer BD FACS Canto II (Immunocytometry Systems, BD, New Jersey). Acquired data were analyzed by the Flow Jo software [21 (link)].

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Tian J., Wang W., Zhu J., Zhuang Y., Qi C., Cai Z., Yan W., Lu W, & Shang A. (2022). Histone Methyltransferase SETDB1 Promotes Immune Evasion in Colorectal Cancer via FOSB-Mediated Downregulation of MicroRNA-22 through BATF3/PD-L1 Pathway. Journal of Immunology Research, 2022, 4012920.

Publication 2022

staining buffer PE-CD3 (130-117-139, Pacific blue-IFN-γ

Buffer Cell Ifn γ T cells

Corresponding Organization : Tongji University

Other organizations : Yancheng Third People's Hospital, Huzhou Central Hospital, Soochow University, Yancheng First People's Hospital, Nantong University

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Variable analysis

independent variables

Treatment of Jurkat T cells with PE-CD3 (130-117-139, 1 : 50, Miltenyi Biotec, Bergisch Gladbach, Germany)

dependent variables

Detection of Pacific blue-IFN-γ (BioLegend, San Diego, CA, #505817, rat, 1 : 50) in Jurkat T cells using flow cytometer BD FACS Canto II (Immunocytometry Systems, BD, New Jersey)

control variables

Dispersion of cells into a single-cell suspension and resuspension in staining buffer (BD, New Jersey, USA)
Fixing and permeabilization of Jurkat T cells

positive controls

No positive controls were explicitly mentioned

negative controls

No negative controls were explicitly mentioned

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