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PHA/IL-2 preactivated cells were re-stimulated with 5 ng/mL IL-12 for 15 hours or 20 ng/mL phorbol 12-myristate-13-acetate (PMA) and 1 ng/mL of the calcium-ionophore A23187 (both Sigma Aldrich) for 6 hours, in the presence of GolgiPlug (BD Biosciences) the last 12 and 5 hours, respectively. Cells were permeabilised with saponin and stained with an anti-IFN-γ (B27) or an isotype control (MOPC-21, both BD Biosciences) mAb. IL-12-specific production of IFN-γ was determined by subtracting the frequency of IFN-γ+ cells in unstimulated cells.
Copyright and License information: ©2018 Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.
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