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KEGG signal transduction pathways [ 21] were analyzed for overrepresentation in the triple negative subtype using Fisher's exact test and pathway expression was determined using the mean of normalized pathway components.
Since the raw pathway activity scores (PA) generated from PAPi for different pathways are not comparable, they are further linearly normalized to 0 1, as normalized pathway activity scores (PAN), before aggregation.
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After obtaining the normalized pathway-gene and pathway-gene pair relationships, we are able to analyze the comprehensiveness and agreement among the different pathway databases on different aspects.
We turn complicated and incompatible xml data formats and inconsistent gene and gene relationship representations from different source databases into normalized and unified pathway-gene and pathway-gene pair relationships neatly recorded in simple tab-delimited text format and MySQL tables, which facilitates convenient automatic computation and large-scale referencing in many related studies.
Numbers of normalized reads/library for each pathway were extracted from the local database; non-biologically meaningful pathways (e.g. human disease or other organism-specific pathways) were removed, and the data filtered for minimum KEGG terms/pathway (threshold ≥ 5) and total reads/pathway (≥ 200).
This method, given a set of pathways and normalized gene expression data, allows the transformation of the data into pathway enrichment scores (a measure of the state of each pathway), generating a pathway expression matrix.
The entire distribution of the ESTs in each pathway is normalized to 1 or 100%.
For correlation with biochemical pathways, we computed for each pair of community-pathway the community-wise and pathway-wise specificities, defined as the number of reactions shared by both the community and pathway and normalized by the size of the community and size of the pathway respectively.
While high-dose supplemental CoQ10 therapy normalized expression of several pathways in Pdss2 missense mutants, its effect was less pronounced than that of probucol (Fig 4).
The final evaluation scores are obtained by computing a running-sum statistic across the EnrichNet and ORA ranks for all gene sets from each reference collection, with positive score contributions for benchmark pathways and negative contributions for all other pathways, using the normalized Kolmogorov Smirnov test as defined in Mootha et al. (2003).
INR, international normalized ratio; TFPI, Tissue factor pathway inhibitor.
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