Hyperactivation of cytokines and intracellular mediators in TLR2-stimulated MOLF/Ei macrophages. (A) Peritoneal macrophages from C57BL/6J and MOLF/Ei mice were stimulated with 2 µg/ml LTA for the indicated times. Cytokine mRNA was quantified by real-time PCR. Data represent mean ± range from duplicate wells. One of at least three representative experiments is shown. (B) MEFs were prepared from MOLF/Ei and C57BL/6J mice and stimulated with 100 ng/ml human IL-1β or TNF-α for the indicated times. IL-6 mRNA was quantified using real-time PCR. Data are shown as mean ± range from duplicate wells, representative of two independent experiments. (C–E) BMDMs from each strain were stimulated with 2 µg/ml LTA for the indicated times. Cytoplasmic protein lysates were analyzed by Western blot analysis for MAPK (C) and NF-κB (D) pathway activity or IRAK-1 degradation (E). Data are representative of three (C and D) or two (E) independent experiments.