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Intracellular cytokine staining and flow cytometry

YK Youra Kang
MT Maheshwor Timilshina
TN Tae-gyu Nam
BJ Byeong-Seon Jeong
JC Jae-Hoon Chang
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CD4+ T cells were collected and re-stimulated for 4 h with phorbol 12-myristate 13-acetate (PMA) (50 ng/ml; Sigma) and ionomycin (750 ng/ml; Calbiochem, La Jolla, CA, USA) with GolgiStop (BD Biosciences). Cells were stained with anti-mouse CD4-FITC (GK1.5; BioLegend, San Diego, CA, USA), anti-mouse B220-PE/Cy7 (RA3-6B2; BioLegend), anti-mouse CD3 ε-APC (145-2C11; BioLegend), anti-mouse CD8a-PE/Cy7 (53-6.7; BioLegend), anti-mouse IFN- γ-PE (XMG1.2; BioLegend), and anti-mouse IL-17A-APC (TC11-18H10.1; BioLegend) according to the manufacturer’s instructions. Data were obtained with a FACSVerse (BD Immunocytometry System, San Jose, CA, USA) and analyzed using FlowJo software.

Copyright and License information: The Author(s) ©2017
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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