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. 2009 Sep;128(1 Pt 2):e325–e334. doi: 10.1111/j.1365-2567.2008.02966.x

Figure 4.

Figure 4

CCL5 induces tumour necrosis factor-α (TNF-α) secretion via MEK-ERK activation and disulfiram (DSF) inhibits release of TNF-α from RAW264.7 cells. (a, c) RAW264.7 cells were pretreated with 5 μg/ml neutralizing anti-CCR5 antibody or isotype-matched immunoglobulin G (IgG). (b, d) Cells were pretreated with PD98059 (50 μm), wortmannin (100 nm) and DSF (0·1–0·3 μm) or medium with an equal volume of drug solvent. After 30 min, CCL5 (50 nm) was added for another 6 hr. TNF-α in the supernatants was determined by enzyme-linked immunosorbent assay (a, b) and by biological activity (c, d), as described in the Materials and methods. Results are expressed as the mean ± SD of three independent experiments. *P< 0·05 compared with medium, †P< 0·05 compared with CCL5 and isotype-matched IgG, #P< 0·05 compared with CCL5 (50 nm) stimulation.