FIGURE 4.

MDP-induced degradation of Nod2 and Rip2 is specific to MDP stimulation. A, RAW264.7 cells were pretreated with LPS (0.1 ng/ml) or IFN-γ (100 units/ml) for 6 h and then stimulated with LPS, CpG-B, or MDP for the indicated times. B, wild-type, Nod2-deficient, or Rip2-deficient BMDM were pretreated with LPS (0.2 ng/ml) for 6 h and then stimulated with MDP (100 μg/ml) for the indicated times. C, RAW264.7 cells were pretreated with LPS (1 ng/ml) for 6 h and then incubated with MDP (MDP-ld) or biologically inactive MDP-ll (100 μg/ml) for 3 h. D, wild-type or MyD88-deficient BMDM were stimulated with LPS for 6 h and then infected with heat-killed B. subtilis (5 × 10⁶ or 20 × 10⁶ cells/ml) in the presence of cycloheximide (CHX; 20 μg/ml) for 4 h. Cell extracts were subjected to Western blot analysis for Nod2, Rip2, and actin. Data are representative of three independent experiments with similar results. S.E., short exposure; L.E., long exposure; n.s., nonspecific.