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Triplicate samples were run on a LightCycler 480 Real time PCR system (Roche Diagnostics, Lewes, UK) and expression ratios were calculated using the ddCT method.18Gene expression profilingTotal RNA (100 for each sample) was amplified and labeled with the 3 IVT Express Kit (Affymetrix, Santa Clara, CA, USA), and hybridized to Affymetrix Human Genome U133 Plus 2.0 GeneChips, as described previously.19, 20 CEL files were pre processed using Robust Multi chip Average and data analysis was performed using GeneSpring 7.3.1 (Agilent, Santa Clara, CA, USA).
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