Journal: The Journal of Biological Chemistry
Article Title: Distinct Roles for Nod2 Protein and Autocrine Interleukin-1? in Muramyl Dipeptide-induced Mitogen-activated Protein Kinase Activation and Cytokine Secretion in Human Macrophages *
Figure Lengend Snippet: Decreased MAPK activation upon Nod2 signaling in the absence of the IL-1β autocrine loop persists over time. Human MDMs from healthy controls ( n = 6–8) were stimulated with 100 μg/ml MDP, 10 ng/ml IL-1β, or 100 μg/ml MDP with 0.5 μg/ml IL-1Ra and 1 μg/ml anti-IL-1β antibody for 10, 30, or 60 min and analyzed by flow cytometry for the expression of phospho-JNK, phospho-ERK, or phospho-p38. A , shown are representative flow cytometry plots with the indicated mean fluorescence intensity values. Stimulated cells stained with isotype controls are shown. B , summarized data are represented as the -fold phospho-MAPK induction normalized to untreated cells + S.E. ( error bars ). *, p
Article Snippet: For IL-1β blockade, IL-1R antagonist (IL-1Ra) (GenScript, Piscataway, NJ) and/or anti-IL-1β-blocking antibody (R & D Systems, Minneapolis, MN) was added 1 h prior to treatments with MDP (Bachem, King of Prussia, PA), IL-1β (eBioscience, San Diego, CA), lipid A (Peptides International, Louisville, KY), poly(I:C), CpG DNA (Calbiochem), TriDAP, flagellin, or CL097 (Invivogen).
Techniques: Activation Assay, Flow Cytometry, Cytometry, Expressing, Fluorescence, Staining