This predicted pathway alteration was validated with an animal model of heart failure (HF).
Sequence alterations were validated with a second independent PCR.
KCNQ1OT1 and H19 alterations were validated by methylation specific multiplex ligation-dependent probe amplification (MS-MLPA) using SALSA ME030; and SNRPN alterations were validated with SALSA ME028 (MRC-Holland, Amsterdam, Netherlands).
The identified miRNA expression alterations were validated by real-time PCR, and tissue distribution of the miRNAs was visualized by in situ hybridization.
The array CGH analysis for copy number alterations was validated by FISH.
We further revealed that seven of nine specific genetic alterations that were validated in univariate analysis were enriched in ccB samples with 72% of samples harbouring at least one and up to six of these.
Genomic copy number alterations of selected regions were validated using a relative quantitative (Q -PCR assay.
Using Affymetrix 250K Sty microarrays, we demonstrated improved signal-to-noise ratio and detected significant copy number alterations using the VN algorithm that were validated by independent PCR analysis of the target CNA regions.
Alterations in master regulators and gene targets were validated in PDGF-stimulated smooth muscle cells in vitro and in a model of bladder injury in vivo.
These signatures were based on the combination of clinical data and copy number alterations of a limited number of genes, and were validated in independent series.
