Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-18 induces monocyte chemotactic protein-1 production in macrophages through the phosphatidylinositol 3-kinase/Akt and MEK/ERK1/2 pathways.

doi: 10.4049/jimmunol.175.12.8280

Figure Lengend Snippet: FIGURE 1. IL-18 induces MCP-1 mRNA expression and MCP-1 secretion from murine macrophages. Purified peritoneal macrophages from C57BL/6J mice were treated with medium alone (M) or the indicated concentrations of IL-18 for 6 h for measurement of MCP-1 mRNA (A) or 48 h for measurement of MCP-1 secretion (B). A, Total RNA was isolated and RT-PCR was performed. HPRT was amplified as an internal control. SM, 100-bp DNA ladder. B, Culture supernatant was harvested, and ELISA was performed using a murine MCP-1 ELISA kit. C, Purified macrophages from C57BL/6 mice were treated with IL-18 (5.6 nM) in the presence of the indicated concentrations of anti-IL-18 Ab (IL-18 Ab) or control rat IgG1. Culture supernatant was harvested, and the production of MCP-1 was determined by ELISA. D, Purified macrophages from C57BL/6J mice were treated with medium alone or IL-18 (5.6 nM) for 48 h. Culture supernatant was harvested and placed in the lower chamber of a double-chamber with () or without () anti-MCP-1 blocking Ab (5 g/ml). Freshly isolated monocytes (3 105 cells) were added to the upper chamber, and the number of cells migrating to the lower chamber was counted. Data are shown as the mean SD of triplicate cultures. Data represent at least five (A, B) or three (C, D) independent experiments. , p 0.03; , p 0.00002; , p 0.0004.

Article Snippet: Recombinant mouse IL-18, rat IgG1 isotype, anti-IL-18 Ab, and anti-mouse JE/CCL2blocking Ab were purchased from R&D Systems.

Techniques: Expressing, Isolation, Reverse Transcription Polymerase Chain Reaction, Control, Enzyme-linked Immunosorbent Assay, Blocking Assay