Figure 7.
Effects of anti-CCR5 antibody, PD98059, wortmannin and disulfiram (DSF), on CCL5-induced nuclear factor-κB (NF-κB) transcriptional activity in RAW264.7 cells. RAW264.7 cells (2 × 105) were cotransfected with the mixture of pGL3.5XκB-luciferase, and pRL-TK-Renilla-luciferase for 24 hr. (a) RAW264.7 cells were treated with 5 μg/ml neutralizing anti-CCR5 antibody or isotype-matched immunoglobulin G (IgG). (b) Cells were treated with PD98059 (50 μm), wortmannin (100 nm) and DSF (0·1–0·3 μm) or medium with equal volume of drug solvent; 30 min later, CCL5 (50 nm) was added for 6 hr. The NF-κB luciferase activity was measured using Dual-Luciferase Reporter Assay System and normalized with Renilla luciferase activity. *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.