Figure 5

NF-κB involvement in CS medium stimulation of human macrophages for IL-8 production. a). Cells were left in culture medium or incubated with proteasome inhibitor, MG-132, at 10 μM for 1 hr prior to a 45 minute treatment with CS medium or LPS. Macrophages were lysed to determine IκB-α and phosphorylated IκB-α. b). Macrophages were treated for 30 min with SB 203580 (5 μM) and curcumin (25 μM) prior to CS medium exposure. IL-8 production was quantified using ELISA. c). Cells were left in culture medium or incubated with anti-TLR4 or isotype control prior to a 30 minute exposure to CS or LPS. Nuclear proteins were extracted, subjected to 10 % SDS-PAGE, and blotted with P65 Abs. The result shown is a representative of three experiments conducted with different human monocyte-derived macrophage preparations that had similar results.