Expression data from Jurkat cell line infected with pMIN-ERG+85 vector. Expression data from Jurkat cell line infected with pMIN-ERG+85 vector

Jurkat cells infected with pMIN-ERG+85 express different tagBFP levels. Using flow cytometry sorting, highest and lowest tagBFP cells (10%) were sorted. High tagBFP cells are functionaly have more leukemia stem cell proprties in comparision to low tagBFP cells. Utilizing sorted cells to identifiy differential transcriptomics between two subpopulations We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of up/down-regulated genes that determine High tagBFP cells vs Low tagBFP Overall design: Viral particles were generated by transient transfection of 293T cells using CMVDeltaR8.91 and pMD2G constructs as described elsewhere (Milyavsky et al. 2010). Jurkat leukemia cells were infected by addition of viral supernatant to obtain 10-30% infection rate. Jurkat cells were sorted using BD FACSAria™ Fusion sorter (BD Biosciences), RNA was extracted with the Trizol reagent (Invitrogen) and reverse transcribed with SuperScript III (Invitrogen),RNA from 100,000 or more sorted cells was used for microarray analysis by Human Gene Clariom S Assay (Thermo Fisher Scientific)