GATA-2 expression is confined to HSPCs. (A) HEK-293T cells were transfected with empty vector (EV), or constructs encoding FLAG-tagged GATA-2 or GATA-3 and Myc-tagged PLZF. Twenty-four hours after transfection, GATA-2 or GATA3 were immunoprecipitated and precipitates as well as whole-cell lysates (WCL) were analyzed by western blotting with antibodies, as indicated. (B-D) Bone marrow, cord blood, or PBMCs from healthy volunteers were surface stained with antibodies to lineage and differentiation markers as well as intracellular with antibodies to GATA-2, GATA-3, and PLZF. (B) Barnes-Hut t-distributed stochastic neighbor embedding (t-SNE) analysis of 13-parametric data were performed on gated CD3⁻CD14⁻CD19⁻CD123⁻ cells combined from samples of bone marrow (n = 2) and peripheral blood of CMV seronegative (n = 2) and CMV seronegative (n = 2) from healthy volunteers. Plots show expression of surface and intracellular proteins, as indicated, in the 2-dimensional t-SNE field with relative expression denoted as red when high and blue when absent. (C) Cell density in the t-SNE field for pooled samples from bone marrow (n = 2) and peripheral blood of CMV-seronegative (n = 2) and CMV-seropositive (n = 2) donors, respectively. (D) Contour plots show expression of the indicated markers in bone marrow (CD3⁻CD14⁻CD19⁻CD123⁻) Lin⁻CD34⁺ cells as well as peripheral blood Lin⁻ lymphocytes. In plots depicting CD56 expression, dotted lines discriminate CD56^bright from CD56^dim cells. Plots are from representative donors.