To generate the 454 library, 500 ng of normalized cDNA were used. cDNA was fractionated into small, 300-to 800-basepair fragments and the specific A and B adaptors were ligated to both the 39 and 59 ends of the fragments. The A and B adaptors were domain; PKC: Protein kinase C; PTEN: Phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN; RAIDD: Caspase and RIP adapter with death domain; TNF R1: Tumor necrosis factor receptor 1; TNF-a: Tumor necrosis factor alpha; TRADD: TNF receptor type 1-associated DEATH domain protein; TRAF2: TNF receptor-associated factor 2; TRAIL: TNF-related apoptosis-inducing ligand; TRAIL decoy: Decoy TRAIL receptor without death domain; TRAIL-R: TRAIL receptor. doi:10.1371/journal.pone.0035009.g008 used for purification, amplification, and sequencing steps. One sequencing run was performed on the GS-FLX using Titanium chemistry. 454 Sequencing is based on sequencing-by-synthesis, addition of one nucleotide, or more, complementary to the template strand results in a chemiluminescent signal recorded by the CCD camera within the instrument. The signal strength is proportional to the number of nucleotides incorporated in a single nucleotide flow. All reagents and protocols used were from Roche 454 Life Sciences, USA.