CTR- or RAPA-DC were purified following 7 day culture via positive selection using CD11c beads and then remained unstimulated (Unstim.) or were incubated with 1 μg/ml LPS, 2 μg/ml CpG, or 5 μg/ml agonistic α-CD40 mAb for 20 min, as indicated. (A) MAPK activation was determined by Western blot analysis of DC lysates for phosphorylated and total JNK, p38, and ERK. Blots shown are representative of three experiments with similar results. (B and C) Nuclear extracts from these cells were assessed for DNA binding activity in EMSA using 32P-labeled NF- κB (B) or AP-1 (C) probes. The data shown are representative of at least 2 independent experiments.