Fig. 3

Enhancement of the immunosuppressive function by mTOR inhibition has no involvement with the inflammatory cytokine pathway. a MSCs were pretreated with 10 nM or 100 nM rapamycin (RAPA) followed by treatment with 10 ng/ml tumor necrosis factor alpha (TNF-α) plus 20 ng/ml interferon gamma (IFN-γ) for 1, 3, and 5 days. mRNA expression of the IFN-γ and TNF-α receptors IFNGR1, IFNGR2, TNFR1, and TNFR2 was measured by quantitative RT-PCR. Cells without treatment with rapamycin and inflammatory cytokines were indicated as control. b,c MSCs were treated with 10 ng/ml TNF-α plus 20 ng/ml IFN-γ for the indicated time. The activation of mTOR substrates was determined by Western blot (b, representative data; c, pooled data). β-actin was used as internal control. Data represent mean ± SD of at least three independent experiments