Hepcidin gene expression is increased in airway epithelial cells after exposure to pro-inflammatory cytokines. NHBE cells, grown at ALI, were exposed to LPS (100 μg/ml) + CD14 (250 ng/ml) or to cytomix (TNF-α, IL1-β, and IFN-γ (100 ng/ml, each)) for 1 hr and harvested after 6 hrs (A) or after 2, 6, and 24 hrs to establish a time response curve (B). Additional cells were exposed to individual cytokines, TNF-α, IL1-β, and IFN-γ for 1 hr and harvested after 6 hrs (C) or IFN-γ for 1 hr and harvested after 2, 6, and 24 hrs and compared to time-based controls (D). Total RNA was isolated using RNeasy® and reversed transcribed. Quantitative PCR was performed using TaqMan polymerase. Fluorescence was detected on an ABI Prism 7700 sequence detector (Applied Biosystems). Relative abundance of hepcidin was normalized to 18s and expressed as fold induction over control ± SE and represent at least n = 4. * P < 0.001, #P < 0.05 relative to time-based HBSS control cells.