Figure 3.

Tight junction formation. Images showing staining for zonula occludens-1 (ZO-1; red) tight junctions in static control and rotating ALI cultures (A–F). Note the initial formation of a tight junction network in static cultures and subsequent deterioration by day 21 (A–C) compared to the steady increase in junction density and organization over time in the rotating ALI cultures (D–F). Nuclei are stained with DAPI (blue). Quantitative assessment of cell ZO-1 expression from immunostaining confirmed this trend and is expressed as percentage of cells staining positive, with 95% confidence intervals (G). Expression of ZO-1 at day 14 and day 21 in NHBE cells cultured in rotating ALI culture system compared to static (submerged) controls (H).