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RNA analysis, whole transcript labelling and Affymetrix Exon microarray hybridisation was performed by UCL Genomics (www.genomics.ucl.ac.uk).ac.uk

Sample preparation and microarray hybridisation was performed as described previously,[15] apart from the different postprocessing and prehybridization described above.

Preparation of cDNA SPIA probes for microarray hybridisation, was carried out with varying amounts of total HUVEC RNA (50 ng–10 pg) or vascular endothelial cell samples using NuGEN Technologies (California, USA) WT-Ovation™ FFPE RNA amplification system V2 or WT-Ovation™ One-Direct RNA amplification system according to the manufacturers' instructions.

The cDNA used for the microarray hybridisation was first diluted 10-fold.

RNA labelling and microarray hybridisation was carried out in collaboration with CXR Biosciences, Dundee, UK.

RNA extraction and microarray hybridisation was performed as described in [ 28, 52, 53].

To test the validity of signal quantitation by the Pinus microarray, 60-mer probes were selected and corresponding transcripts in cDNA transcribed from the original total RNA extracts used for microarray hybridisation were quantified by RT-PCR.

Further details on the microarray hybridisation are available at the MRC/CSC/Imperial College Microarray Centre website (http://microarray.csc.mrc.ac.uk).csc.mrc.ac.uk

The 14 samples from the hippocampus used for microarray hybridisation were also analyzed by qPCR analysis.

Quality control, labelling and microarray hybridisation were performed at the Paterson Institute for Cancer Research, Molecular Biology core facility (Manchester, UK).

Microarray hybridisation is not a first-choice technology in current clinical practice because of sample size requirements and cost issues.