Figure 10.

Roflumilast N-oxide reverses the CSE-induced loss in Foxj1 and Dnai2 of ciliated human bronchial epithelial cells. Differentiated human bronchial epithelial cells (D-BECs) were pre-incubated with roflumilast N-oxide at 2 nM or 1 µM, or vehicle for 30 min before being exposed to CSE at 10% for 3 days. Culture medium with fresh CSE and roflumilast N-oxide was replaced every 24 h. Then, the levels of Foxj1 (A) and Dnai2 (B) mRNA transcripts (upper panels) and protein (lower panels) were determined by quantitative real-time RT-PCR or densitometric evaluation of Western blots, respectively. A representative Western blot illustrating the corresponding incubations is depicted as well. Results are expressed relative to control levels (= 1 for mRNA, = 100% for densitometric protein analyses) and shown as the means ± SEM of cultures from two donors and three experiments per donor. *P < 0.05 versus control, ^#P < 0.05 versus CSE.