Illumina HiSeq 2000 paired end sequencing; Origins of MLL cancer stem cells govern leukemic self-renewal mediated by Î˛-catenin and Hoxa9
Origins of MLL cancer stem cells govern leukemic self-renewal mediated by Î˛-catenin and Hoxa9
Ctnnb1fl/fl mice were crossed with Rosa26-CreER Rosa26-YFP mice to generate Ctnnb1fl/fl RosaCre-ER RosaYFP mice. These mice were crossed with Hoxa9-/- knockout mice to generate Hoxa9-/- Ctnnb1fl/fl RosaCre-ER RosaYFP mice. Compound homozygous animals were used for experiments. For leukemia development experiments, C57BL/6 or SJL mice were given 11Gy total body Îł-irradiation and injected with up to 500,000 test cells mixed with 200,000 C57BL/6 or SJL bone marrow nuclear cells. LSK (lin-, Sca-1+, c-Kit+) and GMP (lin-, Sca-1-, c-Kit+, CD34+, CD16/32lo) were isolated from lineage negative cells after lineage (Gr-1, Mac-1, B220, Ter119, CD3e, CD4, CD8) depletion using a BD FACS ARIA cell sorter.  300ng to 1ug of mIRVANA (Ambion) isolated total RNA was used for RNA-Seq library preparation using TruSeq Stranded Total RNA kit (Illumina).
Experimental Factor: hematopoietic stem cell
cell type
Experimental Factor: wild type genotype
genotype