Fig. 3.

Changes in dual NADPH oxidase-putative heme peroxidase (DUOX) mRNA and protein expression after treatment with ATRA. HBE1 cells and primary TBE cells were grown in ALI culture conditions with or without 30 nM ATRA for 7 days followed by harvest for either RNA or protein. A: quantitative real-time PCR (qRT-PCR) analysis of DUOX1 and DUOX2 mRNA expression from HBE1 cells with or without ATRA. ***P = 0.0004 (by ANOVA). B: qRT-PCR analysis of DUOX1 and DUOX2 mRNA expression from primary TBE cells with or without ATRA. **P = 0.0073 (by ANOVA). Data are expressed as fold induction of the gene of interest normalized to β-actin. C: a total of 30 μg of protein from each sample was loaded onto a 7% SDS-PAGE gel, and Western blotting for DUOX was performed using anti-DUOX antibody as described in materials and methods. The membrane was stripped, and Western blotting for β-actin was performed to ensure equal protein loading. Arrow identifies predicted molecular mass of DUOX protein (kDa). D: band intensity was measured using MultiGauge v2.3 software (Fujifilm), and the ratio of DUOX band intensity to β-actin band intensity was calculated. Results are representative of at least 3 separate experiments.