Rapamycin (Rap) mediates proinflammatory effects via activation of JNK and inhibition of TSC2. (A) Human monocytes or (B) THP-1 cells were incubated with 25nM dexamethasone (Dex), 100nM Rap, and 100 ng/mL LPS for 30 minutes as indicated. Whole-cell lysates were analyzed by immunoblot. (C) Monocytes were treated with 20μM SP600125, 100nM Rap, or LPS as indicated. IL-12p40 in the supernatants was determined after 20 hours. Data are mean ± SEM (n = 4). *P < .05. (D) Equal amounts of cytoplasmic and nuclear fractions of THP-1 cells stimulated as indicated for 30 minutes were analyzed by immunoblotting. (E) Monocytes were treated as indicated, and IL-12p40 was analyzed in the supernatants after 20 hours. *P < .05. **P < .01. n.s. indicates not significant. (F) THP-1 cells were transfected with TSC2 or control (Ctrl) siRNA for 72 hours and then stimulated with LPS and/or 25nM Dex as indicated for 24 hours. Whole-cell lysates were analyzed by immunoblot. (G) Monocytes were stimulated as indicated for 24 hours. Whole-cell lysates were analyzed by immunoblot.