Figure 4

Effects NF- κ B on SPLUNC1 production in normal human bronchial epithelial cells (NHBE). NHBE were cultured under air-liquid interface (ALI) conditions. At day 10 of ALI culture, cells were treated with an NF-κB p65 inhibitor helenalin (10 μM) for 2 hrs, followed by Mycoplasma pneumoniae (Mp, 10 cfu/cell) infection for 48 hrs. (A) Helenalin inhibited Mp-induced SPLUNC1 protein at the apical surface of NHBE; (B) NF-κB p65 activity was measured in the extracted nuclear proteins of NHBE using an ELISA-based assay (Active Motif, Carlsbad, CA). Mp significantly increased NF-κB p65 activity, which tended (p = 0.07) to be decreased by helenalin. Data are expressed as means ± SEM (n = 3 replicates).