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. Author manuscript; available in PMC: 2015 Mar 15.
Published in final edited form as: J Immunol. 2014 Aug 8;193(6):2821–2830. doi: 10.4049/jimmunol.1302498

FIGURE 3.

FIGURE 3

mTOR inhibitors attenuate LPS-mediated NOS2 expression and NO production in GM-DC cultures. (A) GM-DCs were either left unstimulated or activated with LPS in the presence or absence of the mTOR inhibitors RAP or KU and the kinetics of nitrite accumulation were measured by Griess reaction daily for the first 72 h after GM-DC stimulation. (B) NOS2 mRNA levels from three independent experiments were analyzed by quantitative RT-PCR 4 h after LPS activation in the presence or absence of RAP. Relative gene expression levels are compared with LPS treatment group. (C) GM-DCs were treated as in (A) for 24 h and analyzed for intracellular iNOS protein expression by FACS. FACS plots are gated on CD11c+ cells. (D) GM-DCs were treated as in (A) and the percentages of iNOS+ GM-DCs (left) and the mean fluorescence intensity (MFI) of iNOS+ GM-DCs (right) were quantified by FACS. All graphs in this figure represent mean values ± SD of at least three independent experiments. *p < 0.05.