Subsequently, the viral load was measured by Cobas Taqman Real time PCR (Roche Molecular Systems, USA) according to the manufacturer's instructions.
Cells were transfected with cDNA encoding human full-length Nox4, full-length Nox2, the Nox2/4 chimera, the Nox4/2 chimera in vector pcDNA3, or empty pcDNA3 with and without co-expression of regulatory subunits (human Rac1G12 V, p47 phox, and p67 phox ), using the FuGene 6 (Roche Molecular Biochemicals) transfection system according to the manufacturer's instructions.
Each TP interval was amplified using the GeneAmp® XL PCR Kit (Roche Molecular Systems, Branchburg, NJ, USA) according to manufacturer's recommended protocol with two reagent mixes.
Genotyping was performed at the Department of Clinical Chemistry of the Erasmus MC (Rotterdam, The Netherlands) using the AmpliChip CYP450 test (Roche Molecular Systems, Alameda, CA, USA) according to the manufacturer's instruction.
The samples were analysed by COBAS AMPLICOR CMV Monitor test (Roche Molecular System, Branchburg, NJ) according to the manufacturer's recommendations.
Analysis of the relative molecular weight of Azotobacter PO was accomplished indirectly in a non-denaturing system according to [Ferguson 1964] with crude extract preparations.
For separation of low molecular weight polypeptides, SDS-PAGE was carried out using a Tris-Tricine buffer system according to the protocol of Schagger and von Jagow [ 22].
The molecular weight distribution was determined by gel permeation chromatography on a TSKgel G2000SWXL column (7.8 mm × 300 mm i.d., 5 µm) with a HPLC system according to the method of Gu et al. [61].
Hybridisation with PCR-amplified probe of the whole GFP gene, labelled with DIG-dUTP (Roche Molecular Systems, Inc., Mannheim, Germany), was done according to manufacturer's instructions.
