Extended Data Fig. 2: Single cell gene expression analysis and mFISH analysis.
a, Representative plots describing the flow cytometry sorting approach used to collect CD34 + /CD38-/CD19 + cells. DAPI and forward scatter were used to identify viable cells (gate p1). To ensure that no dying cells were sorted, the p1 gate was further refined based on cells staining with the cell viability dye carboxyfluorescein succinimidyl ester (CFSE - FITC channel) (gate p2 – used to sort bulk cells). Positive vs negative combinations of specific antibodies were used to identify the populations of interest: CD34 + /CD38-/CD19 + were collected from gate p4. b, Representative heat map from a FLEXsix single-cell Q-PCR transcription panel showing the expression level of selected relevant transcripts in 11 single-cells sorted from the same patient as negative for CD38 expression. UBC is polyubiquitin-C used as a housekeeping gene control, CB8 is a single normal cord blood control cell. c and d, Phylogenetic trees showing the subclonal architectures of Pt10 and Pt11 bulk and CD34+/CD19+/CD38-/low sorted cells.
