Fig. 1

Overview of the experimental strategy. CD34+ cells from healthy, mobilized apheresis donors were immunostained with a 10-fluorochrome panel and single cells were index-sorted into 96-well PCR plates for multiplex qRT-PCR analysis using the Fluidigm Biomark platform. MEP subpopulations were identified by principal component analysis (PCA) and correlated with the original index sorting data and mRNA levels of surface antigens. Identified cellular subsets were validated transcriptionally at the population level and functionally in single-cell differentiation assays. Finally, the cells were ordered in pseudotime to assess differentiation trajectories which were then further validated in functional assays. FACS, fluorescence-activated cell sorting; IF, immunofluorescence; qRT-PCR, quantitative real-time polymerase chain reaction