Nf κb

Peritoneal cells were immediately suspended in RPMI-1640 medium supplemented with 10% FBS, 0.05 mg/ml streptomycin, and 50 U/ml penicillin, and were seeded onto 6-cm dishes (4 × 10⁶ cells per 2 ml/dish). After incubation for 1 h at 37°C in 5% CO₂ to allow adhesion, the medium was discarded to remove nonadherent cells. Adherent macrophages were immediately harvested after washing the dishes with PBS twice for immunoblotting with antibodies to pentraxin-3, MARCO (Bioss), arginase-1, MCP-1, JNK (GeneTex), COX-2 (Cayman Chemical, Ann Arbor, Michigan), NF-κB (Aviva Systems Biology), phosphorylated ERK1/2 (Cell Signaling Technology), or β-actin (Sigma) 19 (link), 22 (link).

Aliquots of 20 μg of protein extracts from HMDMs and HASMCs were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subjected to immunoblotting with antibodies for TSG-6 (ProteinTech Group, Chicago, Illinois), CD36 (R&D Systems), CD68, ACAT1 (Santa Cruz Biotechnology, Santa Cruz, California), ABCA1, collagen-1 (Novus Biologicals, Littleton, Colorado), collagen-3, fibronectin, JNK, α-tubulin, MMP-2 (GeneTex, Irvine, California), MMP-9 (EnoGene, Atlanta, Georgia), TIMP-2 (Abbiotec, San Diego, California), elastin, MARCO (Bioss, Woburn, Massachusetts), MRC1 (LifeSpan BioSciences, Seattle, Washington), phosphoinositide 3-kinase (PI3K), Raf-1 (Abcam, Tokyo, Japan), c-Src (Bioworld Technology, St. Louis Park, Minnesota), phosphorylated ERK1/2 (Cell Signaling Technology, Tokyo, Japan), NF-κB (Aviva Systems Biology, San Diego, California), or β-actin (Sigma) 19 (link), 20 (link), 21 (link), 22 (link), 23 (link).