SHIP is induced via the MyD88 pathway and is responsible for CpG-induced tolerance and cross-tolerance between CpG and LPS. (A) SHIP^+/+ (□) and SHIP^−/− (■) BMmφs were treated with the indicated concentrations of CpG, dsRNA, or LPS and 24 hours cell supernatants assessed for TNF-α (top panels) and IL-6 (bottom panels) by ELISA. Results shown are the mean plus or minus SEM of 3 independent experiments assayed in duplicate. *P < .01 compared with SHIP^+/+. NS indicates not significant. (B) SHIP^+/+ (□) and SHIP^−/− (■) BMmφs were pretreated (pre) or not (0) with 30 nM CpG, 5 μg/mL dsRNA, or 10 ng/mL LPS for 24 hours and then challenged with either 30 nM CpG or 10 ng/mL LPS. At 24 hours, supernatants were assessed for TNF-α and IL-6. Unstimulated cells released less than 50 pg/mL TNF-α or IL-6, and these levels were subtracted. Results shown are the mean plus or minus SEM of 3 independent experiments assayed in duplicate. *P < .02 compared with untolerized cells. **P < .02 compared with untolerized cells. NS indicates not significant. (C) MyD88^+/+ or MyD88^−/− BMmφs were treated or not (0) with 20 ng/mL TGF-β for 8 hours or 100 ng/mL LPS for 24 hours. Whole cell lysates were subjected to immunoblot analyses for SHIP or Shc. Results are typical of 3 independent experiments. (D) MyD88^+/+ (□) or MyD88^−/− (■) BMmφs were treated with CpG or LPS for 24 hours and cell supernatants assessed for TGF-β by ELISA. TGF-β levels detected from unstimulated cells were less than 20 pg/mL and were subtracted. Results are the mean plus or minus SEM for 3 independent experiments assayed in duplicate. *P < .001 compared with MyD88^+/+ cells.