Fig. 2

MSCs’ suppressive effect is diminished in the absence of the TNFα/TNFR2 signaling pathway. Activated CFSE⁺ CD4⁺ or CD8⁺ effector T cells were co-cultured with MSCs WT or TNFR2 KO in different MSC/T cell ratios (n = 6). Proliferation of CD4⁺ T cells (a) and CD8⁺ T cells (b) was measured by flow cytometry. The first bar represents the unstimulated T cells alone (n = 6), the second bar represents the bead-stimulated T cells alone in RPMI (n = 8), and the third bar is for the stimulated T cells alone in 50% RPMI+50% MEMα (n = 6). All data are collected from 3 different experiments. c A flow cytometry representative of proliferation assay at a 1:4 MSC/T cell ratio. Non-stimulated T cells have a single peak represented in blue. Upon stimulation, T cells alone or in co-culture with MSCs start to proliferate, and one can see less intensity of CFSE fluorescence. Data are represented as mean value ± SEM. One-way ANOVA analysis was performed to generate P values. ns, non-significant; *P < .05, **P < .01, ***P < .001, ****P < .0001. b Anti-CD3 and anti-CD28 activation beads