PMA synergizes with NTHi to induce MUC5AC expression in human epithelial cell.

<p>(<b>A</b>) PMA synergistically enhanced NTHi-induced MUC5AC expression at the mRNA level in human epithelial HM3 cell, as assessed by performing RT-PCR (left panel) and real-time quantitative PCR (Q-PCR) analysis (right panel). Cyclophilin was used as a control for amount of RNA used in each reaction. (<b>B</b>) PMA synergized with NTHi to induce MUC5AC expression at the transcriptional level in human epithelial cells, as assessed by MUC5AC-dependent promoter Luciferase assay. (<b>C</b>) Synergistic induction of MUC5AC expression by NTHi and PMA was also observed in HM3 cells stably transfected with pMUC5AC 3.7 kb-luc. (<b>D</b>) PMA synergizes with NTHi to induce MUC5AC transcription in a dose-dependent manner and vice versa. (<b>E</b>) PMA synergistically enhanced NTHi-induced MUC5AC expression at the transcriptional level in human airway A549, middle ear HMEEC-1 and primary bronchial epithelial NHBE cells, as assessed by MUC5AC-dependent promoter assays. Values are the means ± S.D. (n = 3). <i>*p<0.05</i> vs. control; <i>**p<0.05</i> vs. NTHi alone. The data shown are representative of three independent experiments. −, absence of; +, presence of; NTHi, nontypeable <i>Haemophilus influenzae</i>.