Figure 3.

The activation of Akt and mTORC1 is important for the commitment to glycolysis and maturation of CD137L-DCs. DCs were differentiated and pre-treated with DMSO or 10 μM LY294002 or 50 nM Rapamycin for 1 h before maturation. Glycolysis stress assays of (A) mature moDCs and (B) mature CD137L-DCs were done with a Seahorse XFe24 Analyzer. (C) Cell surface expression of co-stimulatory and inhibitory molecules was measured by flow cytometry. (D) The secretion of cytokines by DCs was measured by ELISA. Depicted are means ± standard deviations of changes upon addition of 10 μM LY294002 or 50 nM Rapamycin relative to respective DMSO controls (values set at 1) from up to 5 independent experiments with DCs from different donors. ^*p < 0.05, ^**p < 0.01 (two-tailed, one sample student t test). n.d., not detected.