Fig. 4

MSCs trigger Treg accumulation in mice via TGF-β at an early stage. A Scheme of the Co-culture and treatment experimental design. Some mice in the AME group were sacrificed 1 week after AOM application, and naïve primary T cells were extracted from the spleen of mice, and co-cultured with MSCs for 48 h and 72 h respectively. B Flow cytometry analysis of the mean percentage of Th1, Th2 and Treg cells in naïve CD4T cells co-cultured with MSCs (ratio 1:5) for 48 h and 72 h respectively or without MSCs. C Analyze and count separately. We extracted proteins from MSCs co-cultured for 72 h with naïve CD4⁺T. D, E Representative immunoblot bands and histogram of relative expression for the IFN-γ, TNF-α, TGF-β1 IL-4, IL-6, IL-10, IL-12 and IL-17. F Flow cytometry analysis of the mean percentage of Th1, Th2 and Treg cells in naïve CD4⁺T cells co-cultured with TGF-β (10 ng/ml) or without TGF-β. G, H Analyze and count separately. Values are expressed as means ± SD.*P < 0.05, **P < 0.01. MSC mesenchymal stem cells, MT MSCs co-cultured for 72 h with naïve CD4⁺T