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A statistical enrichment analysis of functional categories and pathways were performed using Ingenuity Pathway Analysis system IPA 6.0 (Ingenuity Systems, www.ingenuity.com).com
Given the involvement of INT6 in cell proliferation [16], western blots using a panel of antibodies against proteins involved in the cell cycle and associated signalling pathways were performed using lysates from control and INT6 siRNA transfected MDA-MB-231 cells.
Ortholog assignment and mapping of the contigs to the biological pathways were performed using KEGG automatic annotation server (KAAS).
Enzyme prediction and mapping to KEGG biochemical pathways were performed using BLAST2GO and PRIAM (Claudel-Renard et al. 2003).
Statistical analyses (including correction for multiple hypothesis testing) for identification of overrepresented ontologies, functions and pathways were performed using DAVID (http://david.abcc.ncifcrf.gov) and the Partek Genomics Suite.
The searches for gene networks and pathways were performed using DAVID (the Database for Annotation, Visualisation and Integrated Discovery) [ 32] and HEFalMp (Human Experimental/Functional Mapper) [ 33].
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Further investigation of gene functional pathways was performed using Gene Set Enrichment Analysis (http://www.broad.mit.edu/gsea/) as described67.
To test for genetic associations that might escape individual variant testing due to small effect sizes, pathway analysis of eight DNA repair pathways was performed using hierarchical modeling.We identified three susceptibility DNA repair genes, RAD51B (P < 5.09 × 10 -6)), MSH5 (P < 5.09 × 10 -6)), and BRCA2 (P = 5.70 × 10 -6)).
Enrichment analysis of pathways was performed using Fisher's Exact Test.
Hierarchical average linkage clustering of data for selected pathways was performed using MeV v4.2 [101] with Euclidean and Pearson correlation as distance metrics for completeness and activity scores, respectively.
Analysis of regulated pathways was performed using Genemap software.
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