N were analyzed every single 24 h for eight days applying the BD FACSAriaII. Experiments have been performed in triplicate; the typical ten,000 cells per gate had been recorded and analyzed.Extra filesAdditional file 1: Figure S1. Molecular traits of identified subpopulations. a Left: QRT-PCR of GATA1 and GATA2 in K562 cells measured relative to ACTIN. Error bars represent standard error. Appropriate: Representative FACS analysis of K562 cells stained for GATA1 and GATA2. b Histograms displaying expression (fluorescent intensity) of CD44 (light blue) and CD52 (dark blue) in K562 cells. c Dot plots displaying the gating approach for sorting CD24hi and CD24lo expressing K562 cells. d Expression analysis of GATA1 and GATA2 in CD24 sorted K562 cells measured by qRT-PCR relative to ACTIN. P worth 0.05 (t-test), error bars represent typical error. e Representative FACS evaluation of CD24hi and CD24lo sorted K562 cells, stained immediately after sort for pJUN. Imply fluorescent intensity (MFI) was 152 (high), and 137 (low). (PDF 616 kb) Added file two: Figure S2. Molecular traits of identified subpopulations. a Heatmap in the correlation coefficient of K562 CD24 sorted ATAC-seq samples using Spearman correlation. b UCSC tracks displaying examples of open chromatin in CD24lo (blue) and CD24hi (red) K562 cells. Top: JUN locus, which can be more accessible in CD24lo. Bottom: SOD1 locus, that is equally accessible in each subpopulations. c Bar plot illustrating the distribution of ATAC-seq peaks across genomic areas; promoter proximal (light blue) and distal (dark blue). The distinction in promoter accessibility between CD24hi and CD24lo K562 cells is important (working with Chi-squared), p 0.001. d Gene Ontology terms for accessible chromatin locations in CD24lo cells. e Q-Q plot illustrating the differences inside the overlap of ENCODE DNAse-seq peaks and ATAC-seq peaks of CD24lo and CD24hi populations (shown in Fig. 2g). P values for Wilcox (WX), t-test (TT), and Komorov mirnov (KS). f Enrichment of GATA2 ChIP-seq binding websites in CD24hi and CD24lo K562 ATAC-seq peaks. (PDF 507 kb) Additional file three: Figure S3. Functional characteristics of identified subpopulations.Caspase-3/CASP3, Human (His) a Quantification of EdU incorporation as a measurement of proliferation.PDGF-AA Protein Gene ID Experiments had been completed in triplicate; asterisk indicates significance, calculated working with t-test, p value 0.PMID:26780211 05. b Quantification of apoptosis of K562 cells treated with 1 M imatinib or DMSO control for 24 h. AnnexinV I adverse cells are counted as reside, annexin V-positiveLitzenburger et al. Genome Biology (2017) 18:Page 11 ofInstitute of MIT and Harvard, Cambridge, MA 02142, USA. 5Harvard Society of Fellows, Harvard University, Cambridge, MA 02138, USA. Received: eight July 2016 Accepted: 12 DecemberReferences 1. Buffo A, Rite I, Tripathi P, Lepier A, Colak D, Horn AP, et al. Origin and progeny of reactive gliosis: a source of multipotent cells inside the injured brain. Proc Natl Acad Sci U S A. 2008;105(9):3581. 2. Sirko S, Behrendt G, Johansson PA, Tripathi P, Costa M, Bek S, et al. Reactive glia inside the injured brain acquire stem cell properties in response to sonic hedgehog. [corrected]. Cell Stem Cell. 2013;12(four):4269. 3. Friedmann-Morvinski D, Verma IM. Dedifferentiation and reprogramming: origins of cancer stem cells. EMBO Rep. 2014;15(3):2443. 4. Micalizzi DS, Farabaugh SM, Ford HL. Epithelial-mesenchymal transition in cancer: parallels between normal development and tumor progression. J Mammary Gland Biol Neoplasia. 2010;15(2):1174. five. Barski.