Research Paper Volume 12, Issue 24 pp 24734—24777

Figure 1. Overview of the high-risk and low-risk AML patient cohort and the liquid chromatography tandem mass spectrometry (LC-MS/MS) workflow for the proteome and phosphoproteome analysis. The study included AML cell samples from 15 high-risk and 18 low-risk patients collected at the time of first diagnosis. Patients were classified after cytogenetic and molecular genetic analyses. Low-risk patients were further split into elderly low-risk and younger low-risk patients. AML sample preparation steps for proteome and phosphoproteome analysis included AML cell enrichment by density gradient separation, genetic analyses with classification of patients, cell lysis, addition of the super-SILAC (stable isotope labeling with amino acids in cell culture) mix, filter-aided sample preparation (FASP)-based protein digestion and additional immobilized metal affinity chromatography (IMAC) enrichment of phosphopeptides before data-dependent acquisition (DDA) on the mass spectrometer. *Median age of each patient group or subgroup.