SRA

Purpose: To detect the global effects of Class1A PI3K TKO deletion on the HSC transcriptome. Method: Whole BM cells from donor mice TKO: Pik3calox-lox; Pik3cblox-lox; Pik3cd-/-;Mx1-Cre, d KO: Pik3calox-lox; Pik3cblox-lox;Pik3cd-/- (littermate control) and WT: WT, Mx1-Cre (control) were transplanted into lethally irradiated 6-8 week old B6.SJL mice (N=7 per group). Recipient mice were injected 4 weeks post transplant interperitoneally with pIpC 250µg x2, 48 hours apart to ensure Pik3ca and Pik3cb excision. At 4 weeks post PIPC, BM cells were harvested from femurs, tibiae, ilia and vertebrae by gentle crushing. Donor-derived HSCs (Lin-cKit+Sca1+Flk2-CD48-CD150+) cell populations were sorted into RNA extraction buffer (ARCTURUS PicoPure). cDNA was amplified from total RNA using the Ovation® RNA-Seq System V2. Sequencing was performed using the NGS platform llumina-HiSeq2500/4000. Overall design: Examination of transcriptome differences in hematopoietic stem cells between non-competitively transplanted Pik3ca lox/lox;Pik3cblox/lox;Pik3cd-/-;Mx1-Cre (TKO) vs Pik3calox/lox;Pik3cblox/lox;Pik3cd-/- (dKO) littermate controls vs WT;Mx1-Cre (WT) controls all samples are in three biological replicates. Each of the biological replicates was done in 2 technical replicates.