Fig 2: Comparative functional consequences of CD137-costimulation of human CD8+ T cells provided in cis versus trans.a schematic representation of antibody-coated microbeads used to stimulate CD8+ T cells. b semiquantitative FACS assessment of anti-CD3 and anti-CD137 mAbs coupled into the microbeads was measured by antimouse IgG2a and antimouse IgG1 mAbs staining and detected by FACS. Representative histograms of the geometric mean fluorescence intensity for each condition. c, human primary CD8+ T cells from healthy donors were activated with mAb coated microbeads for 96 h. Representative dot plots of cell-surface CD25 (n = 15) and d, intracellular Bcl-xL (n = 13) expression analysed by flow cytometry in the indicated conditions. Summary data are given as delta (?), the difference of the value between each costimulation condition subtracted from the anti-CD3 condition in each case. e representative histograms of CD25 (n = 15) and f, Bcl-xL (n = 14) expression in CD8+ T cells. Numbers in the FACS histograms represent the geometric mean fluorescence intensity (gMFI). Summary data are given. Concentrations of human IFN? (n = 9) (g) IL-2 (n = 8) (h) and granzyme B (n = 4) (i) in the culture supernatants of the indicated experimental conditions. j, representative histograms of T-bet and k, Eomes intracellular analyzed by flow cytometry at the indicated conditions (n = 12). Summary data are given as the difference of the value between each costimulation condition subtracted from the anti-CD3 value in each case. l, summary data showing the T-bet:Eomes ratio of CD8+ T cells in cis versus trans costimulation (n = 12). m, flow cytometry measurement of Mitotracker (n = 11) and n TMRM (Tetramethylrhodamine, Methyl Ester, Perchlorate) (n = 16) stainings given as geometric mean fluorescence in FACS-gated CD8+ T cells after cis versus trans costimulation. Data are given as mean ± s.e.m. Statistical significance was determined with the paired t test (two-sided) in b, c and k, Wilcoxon (two-sided) in d, e, i, j, l and m, the Friedman test (one-sided) with Dunn’s correction in f, g and h. Source data are provided as Source Data file.