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Alcian blue staining of mucus.
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ALI culture samples were washed in distilled water and permeabilized for 10 min with 100 µL of 1 g/L Triton in PBS stained with Alcian Blue (American Mastertech KTAPAPT kit and then stained with 1 × 10−5 M DAPI in PBS (Molecular Probes D-1306 lot 4991-27) for 15 min. Samples were imaged as aforementioned using Keyence BZ microscope and Z-stack imagines were used for analysis.
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