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The total unigene dataset was further filtered using the Affymetrix probe selection pipeline.
Microarray expression ratios were presented as log-transformed base 2. Gene annotations were obtained through batch BLASTs (BLASTx; E-value < e-10) using the Affymetrix probe set sequences in tandem with sourcing annotations directly from the Affymetrix website.
To investigate the possible gene expression change in our ConA-induced hepatitis model at a very early stage, we performed a gene chip study by using the Affymetrix probe dataset which includes totally 45101 probes.
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In the Uppsala series [27] we could assess the precursor miR-210 using the Affymetrix probes (Affymetrix U133b, 230710_at) and this also showed an inverse relationship of the precursor to ISCU and poor survival (Figure 5B).
We used the Affymetrix probe sets verbatim, but discarded those not included in the Ensembl database for the RAT230 2 chip prior to the statistical analysis, reducing the set of probes from 31,099 to 12,919.
We grouped the 1592 breast cancer patients on the basis of the expression of PSMB7 using the Affymetrix HGU133A probe set 200786_at.
Signal intensity for each gene was calculated using the Affymetrix MAS 5.0 probe level algorithm.
To match the Affymetrix gene expression data used below, the Affymetrix probe set annotation was used, as described above.
The blasn program was used to compare the sequences of ESTs used for the Affymetrix probe design with the sequences of ESTs mapped to the wheat deletion bin map [ 48].
However, as on both these platforms oligonucleotides in a set are identical, we aligned all probes against Ensembl genes, with the version used for the Affymetrix probe sets, to ensure the same genome builds were consistently used (Mouse NCBI m34 and Rat RGSC 3.4).
NKI samples were profiled on the Agilent Human 25 K platform comprised of 24,496 non-control oligonucletoide probes while the other studies were carried out using the Affymetrix HG133A platform, compromised of 22,282 probe sets.
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