Human ht12 expression v 4 bead arrays

Total RNAs from TNFα-stimulated FDCLCs were purified using NucleoSpin RNA (740955, Macherey-Nagel, Düren, Germany). The purity and integrity of the isolated RNAs were evaluated by denaturing gel electrophoresis, via the OD 260/280 ratio, and by analysis on an Agilent 2100 Bioanalyzer (G2939BA, Agilent Technologies, Palo Alto, CA). cRNAs were generated from total RNA extracts using an Ambion Illumina RNA Amplification Kit (AMIL1791, Ambion, Austin, TX) and hybridized to Human HT12 Expression v.4 Bead Arrays in accordance with the manufacturer's instructions (Illumina, Inc., San Diego, CA). Array signals were detected using Amersham fluorolink streptavidin-Cy3 (GE Healthcare Bio-Sciences, Little Chalfont, UK) and an Illumina BeadArray Reader Confocal Scanner.

Total RNA was amplified and purified using the Ambion Illumina RNA amplification kit (Ambion, Austin, TX) to yield biotinylated complementary DNA (cDNA) according to the manufacturer's instruction. Briefly, 550 ng of total RNA was reverse-transcribed to cDNA using deoxythymidine oligomer primer and labeled with biotinylated deoxyribonucleotide triphosphate. Labeled cDNA samples were hybridized to human HT-12 expression v.4 bead arrays, and the signal intensity was detected according to the manufacturer's instruction (Illumina, San Diego, CA). Raw data were processed by log2 transformation and quantile normalization.