Whole-genome gene expression profiling of Pik3cg-depleted mice

We performed whole-genome gene expression profiling in Pik3cg-/- mice and subsequent gene ontology clustering of differentially expressed genes compared to wild type mice, in order to investigate the role of Pik3cg in platelet membrane biogenesis and blood coagulation. Pik3cg-deficient mice were obtained from sources previously described (T. Sasaki et al, 2000, Science 287, 1040-1046), backcrossed onto the C57BL/6J Jax genetic background for eight generations (B6J;129-Pik3cg-tm1Pngr) and then maintained as a closed colony by intercrossing from within the colony (C57BL/6J Jax contribution: 99.6%). Whole blood from three Pik3cg-/- and three C57BL/6J Jax wild type mice (all females, age: 13-16 weeks, Mouse Breeders Diet (Lab Diets 5021-3)) was collected from terminally anaesthetized mice via the retro-orbital sinus. For each RNA extraction, in vitro transcription (IVT) reactions were performed in duplicates.