Fig. 4: Identification of markers to dissect transitional cell states.
From: An immunophenotype-coupled transcriptomic atlas of human hematopoietic progenitors
a, Illustration of the donor cohort and Infinity Flow experimental design. b, Expression of surface markers that could enable purification of committed granulocyte progenitors. c, Proposed gating for C5L2+TSPAN33+ cells in Infinity Flow (top) and representative cytometry sorting (bottom). d, Cytospin morphology of cells sorted based on C5L2+TSPAN33+ expression, Wright–Giemsa staining of low (top left), mid (top right) and high (bottom left) and Quick III staining of high (bottom right). e, Differentiation potential of C5L2+TSPAN33+ sorted cells by c.f.u. assay. Data are shown as mean ± s.d. f, Cell composition in a virtually gated MEP subset by CD133 expression in CITE-seq data; Ba/Ma/Eo, basophil/mast cell/eosinophil; Neu, neutrophil. g, CD71 expression in immunophenotypically defined MEPs by CD133 expression in Infinity Flow. h, MEP subset by CD133 expression in cytometry sorting. i, c.f.u. readout of MEP cells sorted based on CD133 expression from the donor in h (data from the other donor are shown in Extended Data Fig. 9h). Blue brackets highlight the proportion of megakaryocytic/erythroid single and bipotential colonies. j, CD326 and CD235a denote three distinct cell populations in Infinity Flow (top) and cytometry sorting (bottom) that are different in cell size and CD71 expression. k, Morphology of sorted cells based on CD326 and CD235a expression; CD326+CD235a–, proerythroblasts; CD326+CD235a+, basophilic erythroblasts; CD326–CD235a+, (1) reticulocyte, (2) polychromatic erythroblast, (3) orthochromatic erythroblast, (4) basophilic erythroblast, (5) mature RBC, (6) transitioning polychromatic-to-orthochromatic erythroblast, (7) orthochromatic erythroblast and (8) orthochromatic erythroblast. The Infinity Flow object shown is representative of donor WM29. The data from the c.f.u. assays are the results of three replicates from two donors either combined (e) or displayed individually (i). Data in d and k are representative of ten high-power fields (differential count in Extended Data Fig. 9); c.f.u. types: granulocytic (G), monocytic (M), erythroid (E) and megakaryocytic (Mk) cells.
