Instruction of haematopoietic lineage choices, evolution of transcriptional landscapes and cancer stem cell hierarchies derived from an AML1-ETO mouse model
Instruction of haematopoietic lineage choices, evolution of transcriptional landscapes and cancer stem cell hierarchies derived from an AML1-ETO mouse model
C57BL/6 recipients (2 x 5.5 Gy)  were injected with 5 x 105 non-induced R26/AE BM cells and exposed to DOX for ten days For obtaining cells for the following RNA-Seq experiments, 4 x 10^4 L-K+S+IL7Ra-CD34+FcgRII/III+ cells were preparatively sorted using a FACSAria cell sorter (BD). RNA was extracted with the Pico Pure RNA Extraction Kit (Arcturus) according to the manufacturers instructions. The resulting RNA was analysed using an Agilent Bioanalyzer. RNA samples were amplified with Nugen`s Ovation RNA-Seq System and quantified by Qubit measurement. The TruSeq DNA Sample PrepKit (Illumina) was used and the resulting cDNAs were fragmented using AFA technology (Covaris). Subsequently, cDNA fragments were end repaired and 3' adenylated. Illuminas TruSeq adaptors with integrated indices were ligated and the resultant fragments size selected using 2% E-Gels (Invitrogen). Enrichment was done by linear PCR and the resulting fragments were purified using Agencourt AMPure XP magnetic beads.
Experimental Factor: time
10 day
Experimental Factor: disease
indolent myeloproliferative like myeloid leukaemia