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All data was analysed in duplicate with SPSS 15.0 software (SPSS Inc., Chicago, IL, USA).
Four vials were analysed in duplicate with reagents from Riedel-de Haën, Seelze, DE.
All serum samples were analysed in duplicate with each replicate being randomised across days and chips.
Matched pairs were analysed in duplicate with a dye swap for the second microarray to reduce the effects of dye incorporation bias.
For the qPCR analysis, 1/10 and 1/100 dilutions were made in TE (10 mM Tris-0.2 mM EDTA pH 8.0) and analysed in duplicate with the VitB2-UGM qPCR assay as described above.
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Samples and standards were analysed in duplicates with a maximum tolerated coefficient of variation (CV) of 20%.
All analyses were performed in duplicate, with each experiments performed at least twice.
Briefly, quantitative real time RT-PCR analyses were performed in duplicate with an ABI 7900 Fast Sequence Detection System using TaqMan gene expressionn assays and universal PCR master mix according to the manufacturer's specifications (life technologies) [ 15].
Quantitative real time PCR analyses were performed in duplicate with an ABI 7900 Fast Sequence Detection System using TaqMan gene expression assays and universal PCR master mix according to the manufacturer's specifications (Applied Biosystems).
To ascertain analytic quality all standards, controls and samples were analysed in duplicate and all duplicates with a CV >10% were reanalysed.
Serially diluted cDNA samples were analysed in duplicate by qRT-PCR with an ABI 7300 Real Time PCR instrument (Applied Biosystems), using Taqman (ABI Masterr Mix, primers/probes for the gene of interest and for a control gene (GAPDH).
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