Fig. 3

FL-MSCs down-modulate CD4⁺ and CD8+ T convs stronger than BM-MSCs. CD3/CD28 activated CD3⁺CD25⁻ effector T cells were co-cultured with BM-MSCs or FL-MSCs in a fixed 1:5 MSC to T cell ratio. After 1 day, T cells were collected and activation markers (CD25, GITR, TNFR2, and ICOS) were analyzed by flow cytometry. Representative flow cytometry dot plots showing the percentage of CD25, GITR, TNFR2, and ICOS, among CD4⁺Foxp3⁻ and CD8⁺Foxp3⁻T cells from the activated T cells control group (left panel). After delimitating the lymphocyte region by a forward-scatter-area (FSC-A) versus side-scatter area (SSC-A) plot, a CD4 versus Foxp3 plot and a CD8 versus Foxp3 plot were used to gate on CD4⁺Foxp3⁻T convs and CD8⁺Foxp3⁻ T cells, respectively. Frames defined the positive subpopulations for each marker analysis in the CD4⁺Foxp3⁻ and CD8⁺Foxp3⁻ populations. Statistical summary dot-plot graphs showing the percentage or the MFI value of each marker analyzed in CD4⁺Foxp3⁻and CD8⁺Foxp3⁻ T convs (right panel). Each dot represents a measured value collected from 2 different experiments (n = 12 for T cells + beads (black) and n = 9 for BM-MSCs + T cells (red) and FL-MSCs + T cells (blue) groups). For each group of values, horizontal lines represent mean value ± SEM. MFI values have been normalized with T cells + beads control group. One-way ANOVA analysis was performed to generate P values. ns, non-significant; Beads, anti-CD3 and anti-CD28 activation beads; T convs, conventional T cells. *P < .05, **P < .01, ***P < .001, ****P < .0001