Figure 1.

Identification of Hlf as a Candidate Regulator of Early Hematopoiesis

(A) Schematic depiction of the progenitor subsets that underlie the early stages of hematopoiesis.

(B) Heatmap showing the expression levels of TFs displaying 2-fold or higher expression in HSCs and GMLPs compared with downstream progenitor cell types (mean values of three replicate arrays per cell type, except for HSCs and pMegEs, for which six and five replicate arrays were used, respectively). See also Table S1.

(C) Schematic depiction of the different lentiviral constructs used for OP9 stromal co-cultures.

(D) The lineage output relative to controls of B cells, NK cells, and myeloid cells produced from Hlf/Hlf, Hlf/Dbp, Hlf/Tef, Hlf/Nfil3, Hlf, Nfil3, and M33-Hlf transduced GMLPs in OP9 stroma co-cultures (n = 6, 6, 6, 6, 6, 3, 6, and 45 cultures, respectively, from two independent experiments).

(E) B, NK, and myeloid cell generation in OP9 co-culture experiments and T cell generation in OP9-DL1 co-cultures using Hlf-inducible GMLPs cultured in the absence or presence of DOX. Shown are representative FACS plots as well as bar graphs showing the frequency relative to untreated controls of CD19⁺ B cells, NK1.1⁺ NK cells, CD11b⁺ myeloid cells, and CD90⁺ T cells in the cultures from three independent experiments (n = 5 for the B, NK, and myeloid experiments, and n = 9 for the T cell experiments).

Error bars denote SEM. CFU-erythroid, colony-forming unit-erythroid; MkP, megakaryocyte progenitor; pCFU-erythroid, pre-colony-forming unit-erythroid; pGM, pre-granulocyte-monocyte progenitor; pMeg/erythroid, pre-megakaryocyte-erythroid progenitor. See also Figures S1 and S2.