One hundred nanograms of total RNA was amplified and labeled with biotin using the Ambion Message Amp Premier RNA Amplification Kit (Thermo Fisher Scientific, Waltham, MA) following the manufacturer’s protocol initially described by Van Gelder et al. (59). Hybridization with the biotin-labeled RNA, staining, and scanning of the chips followed the prescribed procedure outlined in the Affymetrix technical manual and has been previously described (60). The oligonucleotide probe arrays used were the Human Genome U133 Plus 2.0 Arrays, which contain more than 54,000 probe sets representing more than 47,000 transcripts. The array output files were visually inspected for hybridization artifacts and then analyzed using Affymetrix Expression Console version 1.4 using the MAS 5.0 algorithm, scaling probe sets to an average intensity of 500.

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