Figure 1: PDE4 acts as a key negative regulator for NTHi-induced upregulation of CYLD expression.
From: Inhibition of PDE4B suppresses inflammation by increasing expression of the deubiquitinase CYLD
(a–c) CYLD mRNA expression was measured in (a) human lung A549, (b) human middle ear HMEEC or (c) human primary bronchial NHBE cells pretreated with Rolipram (10 μM) for 1 h and stimulated with NTHi for 5 h. (d) HMEEC cells were pretreated with Rolipram for 1 h, followed by NTHi stimulation, and cell lysates were analysed by immunoblotting with the indicated antibodies (upper panel). CYLD protein expression was quantified from three independent experiments (lower panel). CYLD mRNA expression was measured in (e) lung and (f) middle ear tissues from C57BL/6J mice preinoculated with Rolipram (10 mg/kg) for 2 h and inoculated with NTHi for 9 h. Mouse tissues from (g) lung and (h) middle ear were stained against CYLD after NTHi inoculation, with or without inoculation of Rolipram (magnification × 200 in g; × 400 in h, respectively). Scale bars, 20 μm. (i) CYLD mRNA expression in HMEEC (left panel) or mouse middle ear tissues (right panel) was measured after NTHi treatment, pretreated with or without Roflumilast (1 μM for in vitro, 5 mg/kg for in vivo). Data in a–f and i are mean±s.d. (n=3). *P<0.05. Statistical analysis was performed using Student’s t-test. Data are representative of three or more independent experiments.
