FIG 3.

Growth of influenza viruses in HBE cells treated with nocodazole. (A) Uninfected primary HBE cells grown at an air-liquid interface were treated with DMSO or nocodazole (10 μg/ml) on the apical and basal surfaces for 24 h. Fixed transwells were stained with anti-α-tubulin antibody. Intracellular MT and formation of cilia were determined by z-stack acquisition on a confocal microscope. The front view of the cells is displayed by using a clipping plane in the Imaris software. Scale bars are 5 μm. Single representative cells chosen from the entire coverslip are displayed for clarity. HBE cells were infected with A/CA/07/2009 (H1N1) (B) and A/Perth/16/2009 (H3N2) (C) viruses at 10³ TCID₅₀ per well. At 24 hpi, cells were treated with DMSO(−) or nocodazole (10 μg/ml) on the apical and basal surfaces. Medium containing DMSO and nocodazole was maintained on the apical and basal surfaces for the duration of the experiment. The apical supernatant was collected at 48 hpi for viral titration. This experiment was conducted in triplicate, displayed as each data point, with three different patient cell lines as shown.