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Fig. 3. Ciliated cell expression of PKA fusion constructs in infected and fully differentiated human airway epithelial cells. (A,B) Undifferentiated cells were infected with lentiviruses expressing the PKA subunit fusion proteins from either the CMV promoter or the foxj1 promoter. Dually infected and differentiated cells were stained with anti-acetylated tubulin (ac. tubulin) for cilia with an Alexa Fluor-555-coupled secondary antibody. In cells expressing constructs with (A) the CMV promoter, 10-20% of the cells express either the CFP- or YFP-tagged fusion proteins, of which 50% of cells coexpression both fusion proteins. Both ciliated and non-ciliated cells express the proteins (50% each). Circles indicate fields with little or no cilia signal but with expression of tagged fusion proteins. In cells expressing constructs with (B) the foxj1 promoter, 80-90% of ciliated cells express either the CFP- or YFP-tagged fusion proteins with promoter with almost 100% coexpression. No expression is seen in non-ciliated cells (squares). Bar, 50 µM. Cultures were from a single donor and matched with regard to all culture and infection conditions.

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