Figure 1

Timeline of primary human bronchial epithelial cell air-liquid interface (ALI) cultures. Cells were seeded in transwells in immersed culture for 7 days to allow cell proliferation, and then were shifted to ALI for 10 days to induce mucociliary differentiation. Cigarette smoke extract (CSE) and/or IL-13 treatment began with the ALI condition. At day 10, cells were treated with drug and infected with Mycoplasma pneumoniae (Mp). Apical supernatants were harvested at days 1, 3, and 7 post-infection, and cells were harvested at day 7 for mRNA analysis of gene of interest using real-time RT-PCR. SPLUNC1 = short palate, lung, and nasal epithelial clone 1; and hBD2 = human β-defensin-2.