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. Author manuscript; available in PMC: 2013 Aug 3.
Published in final edited form as: Nat Immunol. 2008 Aug 31;9(10):1157–1164. doi: 10.1038/ni.1645

Figure 3.

Figure 3

TLR-mediated induction of IFN-α in pDCs depends on the mTOR signaling pathway. (a) ELISA of IFN-α in supernatants (above) and immunoblot analysis of mTOR in lysates (below) of purified mouse pDCs (5 × 105) transfected for 5 h with control or mTOR-specific siRNA and then stimulated for 24 h with CpG-A. Below, β-actin serves as a loading control. (b) ELISA of IFN-α in supernatants of pDCs treated with rapamycin or the PI(3)K inhibitor LY294002 (1–25 μM), then stimulated with CpG-A and assessed 24 h later. (c) ELISA of IFN-α in supernatants (above) and immunoblot analysis of p70S6K in lysates (below) of mouse pDCs transfected for 5 h with siRNA pools specific for p70S6K1 (S6K1), p70S6K2 (S6K2) or both (S6K1,2), then stimulated for 24 h with CpG-A. *, P < 0.05. (d) ELISA of IFN-α in supernatants of pDCs isolated from wild-type (WT) and Rps6k1−/−Rps6k2−/− double-knockout (S6K1,2-KO) mouse spleens and stimulated in vitro with CpG-A. Data are representative of three independent experiments (a–c) or two independent experiments with at least two mice per group per experiment (d); error bars, s.e.m. of replicate wells.