Research Paper Volume 12, Issue 13 pp 13716—13739

Figure 4. Cytokine data (IL-2, IL-4, IL-5, IL-9) of pre-stimulated T-cell (24 hours) treated with and without Spermidine (in pg/ml) for 48 hours in cognitive decline patients and controls. Cytokines – IL-2, IL-4, IL-5, IL-9 – were measured by Legend-plex (Biolegend) in culture supernatants after 48 hours of Spermidine treatment. T-cells were pre-stimulated by CD3/CD28 antibodies for 24 hours before polyamine usage. Spermidine was used in the following concentrations: 5μM, 10μM, 100μM, 1000μM, 2000μM. Unstimulated cells (UST) were measured as control. The data sets were not Gaussian-distributed and therefore transformed by R studio orderNorm using the bestNormalize package and analyzed. Raw data (individual data, median + interquantile ranges) (A, B, I, J; E, F, M, N) and transformed data are given (mean with lower and upper confidence limits, C, D, K, L, G, H, O, P). n_control = 12; n_CD-patient = 20 * p<0.05; ** p<0.01; *** p<0.001.