Figure 4

IL-8 activation by acapsular C. neoformans is dependent on NF-κB and AP-1. BEAS-2B cells were transiently transfected in duplicate with 200 ng of luciferase reporter plasmid DNA bearing the wild type IL-8 promoter (-133-luc), or mutations (mut) of its AP-1 (AP-1-mut-luc) or NF-κB (NF-κB-mut-luc) binding sites. After overnight incubation, cells were left untreated (white columns) or stimulated (black columns) for 6 h with acapsular C. neoformans (MOI = 20) (A) or TNF-α (20 ng/ml) (B). Cell lysates were collected and assayed for luciferase activity expressed as relative luciferase units (RLU). Data are shown as the mean ± S.D. Results are representative of three independent experiments.