Kinetic of NFAT-CBR expression.

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Kinetic of NFAT-CBR expression. Kinetic of NFAT-CBR expression. A, Sorted TCR transgenic ML-BLITC CD4+ T cells MH-BLITC CD8+ T cells were plated at 104 cells into flat 96-well plates coated with anti-CD3/28 in the presence of human IL-2. At indicated time points, stimulation was stopped, and CBR signals were measured in triplets. At the same time points, cells from parallel wells were analyzed for CD25+CD69+ cell populations within live T cells, as shown in the second from left graph. Additionally, supernatant from wells was used for detection of murine IL-2 (does not detect human IL-2) and IFNy via ELISA and results plotted in the two rightmost graphs. Pooled data from two independent experiments are shown (n = 6 mice). B, Eighteen hours of T-cell stimulation as in (A). Cells were removed from stimulus, washed twice in excess PBS and cultured in TCM only for the indicated time before wells were analyzed as described for (A). All graphs display mean + SEM. Pooled data from two (for MH-BLITC) and 4 (for ML-BLITC) independent experiments is shown (n = 4–8 mice). Martin Szyska et al. Cancer Immunol Res 2018;6:110-120 ©2018 by American Association for Cancer Research