Figure 3.1.2-C. Detection of human, phosphorylated CFTR protein in 
respiratory epithelial cells after infection with Ad-CFTR. Human bronchial 
brushing cells (freshly recovered during bronchoscopy by cytologic brush 
from normal individuals) and cotton rat tracheal-bronchial brushing cells 
(recovered by cytologic brush from naive cotton rats) were infected in 
vitro with Ad-CFTR or the control Ela deletion mutant virus Ad-dl312 and 
collected 48 hr later (Appendix 1) . As a positive control for expression of 
CFTR following Ad-CFTR infection, 293 cells were infected with Ad-CFTR and 
collected 16 hr later. CFTR protein was evaluated by immunoprecipitation 
with a mouse monoclonal antibody against human CFTR followed by phosphory- 
lation using protein kinase and [ 7 - 32 P]ATP (Cheng et al . , 1990; Bargon et 
al., 1992). Human bronchial brushing cells from 2 normal individuals (indi- 
vidual #1, lanes 1,2; individual #2, lanes 3,4) cotton rat tracheal -bron- 
chial brushings cells (lanes 5,6) and 293 cells (lanes 7,8) were evaluated. 
Shown are control uninfected cells (lanes 1,5), cells infected with Ad- 
dl312 (lane 3,7), and cells infected with Ad-CFTR (lanes 2, 4, 6, 8). The 
expected size range for CFTR protein is indicated. 
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