Extended Data Figure 8: Expression of functional molecules and surface markers during HSC formation.

a, Bar plot of expression of the documented heptad transcription factors in each single cell during HSC formation. b, Bar plot of expression of the definitive HSC-reprogramming factors in each single cell during HSC formation. c, Mean value of expression (FPKM, log₂) of upregulated innate immune/inflammatory genes in pre-HSCs and/or HSCs. d, Bar plot of expression dynamics of selected surface markers in single cells showing different expression patterns in distinct cell types. e, FACS sorting of E10 mouse AGM cells into four subpopulations: CD47⁺ ECs, CD47⁻ ECs, CD47⁺ pre-HSCs (CD47⁺CD31⁺CD41⁺CD45⁻Ter119⁻), and CD47⁻ pre-HSCs (CD47⁻CD31⁺CD41⁺CD45⁻Ter119⁻). Symbols represent the donor chimaerism in peripheral blood at 16 weeks after transplantation of co-cultured cells with the four subpopulations, respectively. f, FACS sorting of c-Kit⁺CD47⁺ and c-Kit⁺CD47⁻ subpopulations from the E11 mouse AGM region. Symbols represent the donor chimaerism in peripheral blood of recipients at 4 and 16 weeks after direct transplantation of c-Kit⁺CD47⁺ and c-Kit⁺CD47⁻ subpopulations. g, Multi-lineage long-term (>16 weeks) repopulation in peripheral blood, bone marrow, spleen, and thymus of recipients transplanted with E11 HSCs (c-Kit⁺CD47⁺), or with 6-day co-cultures initiated from E10 pre-HSCs (CD31⁺CD45⁻Ter119⁻CD41⁺CD47⁺). Donor-derived (CD45.1⁺CD45.2⁺) myeloid (Mac-1⁺/Gr-1⁺), B-lymphoid (B220⁺), and T-lymphoid (CD3⁺ or CD4⁺/CD8⁺) cells are shown in major haematopoietic organs of a representative recipient.

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