Thylation in ETP-ALL in eight consecutive CpG websites across 64 bases. The sequence of the GATA3 amplicon is indicated in the major, when the sequence at the bottom of each panel indicates the dispensation order. The grey boxes highlight peaks resulting from sequential dispensations of C and T from which DNA methylation was calculated (blue boxes). (A) Representative ETP-ALL sample exhibiting high GATA3 DNA methylation. (B) Representative ETP-ALL sample exhibiting low GATA3 DNA methylation. (PNG 226 kb) Further file 6: Table S2. Next generation sequencing comparing the mutational pattern of Hair Inhibitors targets GATA3low (n = 5) and GATA3high ETP-ALL (n = 13). Sanger Sequencing was performed for selected genes in an extended cohort (n = 70) confirming a high rate of FLT3 mutations in GATA3low ETP-ALL. (PNG 47 kb) More file 7: Figure S5. The GATA3low gene expression profile identified situations with ETP-ALL in an independent cohort of T-ALL. Unsupervised hierarchical cluster analysis of diagnostic bone marrow samples from 55 pediatric individuals with T-ALL, working with a set of genes defined by low GATA3 expression in adult T-ALL. Classification of ETP-ALL and non-ETP-ALL was performed based on the ETP gene expression signature published as well as this dataset. (PNG 484 kb) Extra file eight: Table S3. KEGG pathway annotation of DEG in GATA3low versus GATA3high T-ALL patients. Lists of threefold over- and underexpressed genes of GATA3low individuals were uploaded to DAVID Bioinformatics Resources v6.7. Listed are selected KEGG annotated pathway components significantly enriched in GATA3low up- or downregulated DEGs. (PNG 34 kb) Added file 9: Figure S6. Validation from the GATA3low ETP-ALL cluster in adult individuals with T-ALL employing the ETP signature derived from pediatric T-ALL. Unsupervised hierarchical cluster evaluation of diagnostic bone marrow samples from 83 adult sufferers with T-ALL, using a set of genes defined in pediatric T-ALL [20]. Blue = GATA3low ETP-ALL (n = 11), red = GATA3high (n = 72). Individuals assigned to ETP-ALL and “typical” T-ALL are indicated. (PNG 748 kb)Conclusions GATA3 silencing happens in about a single third of adult ETP-ALL sufferers and is associated with GATA3 DNAFransecky et al. Journal of Hematology Oncology (2016) 9:Page 11 ofAdditional file ten: Table S4. KEGG pathway annotation of DEG in PER-117 cells just before versus right after therapy with decitabine. Lists of 1.5-fold over- and underexpressed genes of cells treated with decitabine (five M) have been uploaded to DAVID Bioinformatics Sources v6.7. Listed are selected KEGG annotated pathway elements considerably enriched in cells treated with decitabine. (PNG 61 kb) Acknowledgements This function was supported by a study grant from the Deutsche Krebshilfe (D-Cystine Epigenetic Reader Domain Mildred Scheel Professur) and also the Wilhelm Sander-Stiftung to CD Baldus. Authors’ contributions LF, MN, and CDB developed and supervised the analysis. LF and MN led the data analysis. UK, SH (Heesch), and MM led the strategies development. SS, TH, KS, and NG acquired the data and performed the data evaluation. CS, JOT, SH (Heller), LHM, KI, and LB performed the experimental validation. LF, MN, and CDB prepared the figures. LF and CDB wrote the manuscript. All authors study and authorized the final manuscript. Competing interests The authors declare that they have no competing interests. Author particulars 1 Division of Hematology and Oncology, Charit? University Hospital Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, 12203 Berlin, Germany. 2Department.