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. 2011 Jul 15;286(36):31347–31360. doi: 10.1074/jbc.M111.232413

FIGURE 9.

FIGURE 9.

Cross-talk of NOTCH1-PI3K signaling pathways during TLR2-, NOD1-, and NOD2-triggered maturation of human DCs. A and B, human DCs were transfected with NOTCH1 siRNA at a final concentration of 100 nm. After 72 h, DCs were treated with CTLA-4 (A) or TGF-β (B) for 6 h and subsequently with a combination of TLR and NLR agonists. Activation of PI3K pathway by means of 4EBP1 phosphorylation was addressed by Western blotting. Blots are representative of three independent experiments. C, pretreatment with GSI-I abrogates Rv0754-, C12-iE-DAP-, and MDP-triggered phosphorylation of 4EBP1 under CTLA-4-induced immunosuppression of human DCs. D and E, inhibition of PI3K signaling axis by LY294002 or rapamycin curtails Rv0754-, C12-iE-DAP-, and MDP-triggered maturation of human DCs under CTLA-4-induced (D) or TGF-β-induced (E) immunosuppression. Immunoblot represents three independent experiments, and bar diagrams are representing mean ± S.E. from three independent donors. *, p < 0.05 versus CTLA-4 and MDP-C12-iE-DAP-Rv0754 or TGF-β and MDP-C12-iE-DAP-Rv0754.