Fig. 5.

Exendin-4 (Ex-4) restores the expression of nuclear FOXA2 in primary airway epithelial cells from normal and diseased lungs. a Immunofluorescence analysis of FOXA2 in normal human primary airway epithelial (NHBE) cells exposed to pyocyanin (PCN). NHBE cells were incubated with Ex-4 for 1 hour before the addition of PCN. After 24 hours, NHBE cells were labeled with anti-FOXA2 antibody, and visualized using a secondary antibody conjugated to Alexa Fluor 594 (red). Nuclei were stained with DAPI (blue). Experiments were independently performed three times. Representative images from one experiment is shown. Quantification of fluorescence signal and statistical analysis are presented in the Fig. S7a. b Ex-4 restores the expression of FOXA2 in NHBE cells and in primary airway epithelial cells derived from cystic fibrosis (CF-DHBE), and large and small airways of chronic obstructive pulmonary disease (COPD-DHBE and COPD-DSAE) lungs. Airway cells were pretreated with Ex-4 for 1 hour prior to addition of PCN for 24 hours. Nuclear and cytoplasmic extracts were analyzed for expression of FOXA2. H3 was used as loading control. Experiments were independently performed three times. Representative Western blots from one experiment are shown. Densitometry analysis of changes in the expression of nuclear and cytoplasmic FOXA2 were compared by using Student’s t-test, and presented in the Fig. S7b–d.