Figure 4. The conditioned media from LPS/ATP-stimulated monocytes induces IκBζ expression in an IL-1β/TNFα-dependent, but IL-18-independent manner.

KG-1 cells (10⁶/ml) were incubated with conditioned media from monocytes (10⁶/ml) that were stimulated with LPS (10 ng/ml, 4 h) and ATP (5 mM, last 15 min) for the indicated time points. For selected time points, the cells were incubated with the conditioned media in the presence of IL-1ra (100 µg/ml), IL-18R Ab (10 µg/ml), TNFα Ab (10 µg/ml), or different combinations of these neutralizing agents. Protein-matched total cell extracts were analyzed by Western blotting using anti-serum against IκBζ and actin Ab. Results are representative of 3 separate experiments.