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Early hematopoietic lineage restrictions directed by Ikaros

Abstract

Ikaros is expressed in early hematopoietic progenitors and is required for lymphoid differentiation. In the absence of Ikaros, there is a lack of markers defining fate restriction along lympho-myeloid pathways, but it is unclear whether formation of specific progenitors or expression of their markers is affected. Here we use a reporter based on Ikaros regulatory elements to separate early progenitors in wild-type and Ikaros-null mice. We found previously undetected Ikaros-null lympho-myeloid progenitors lacking the receptor tyrosine kinase Flt3 that were capable of myeloid but not lymphoid differentiation. In contrast, lack of Ikaros in the common myeloid progenitor resulted in increased formation of erythro-megakaryocytes at the expense of myeloid progenitors. Using this approach, we identify previously unknown pivotal functions for Ikaros in distinct fate 'decisions' in the early hematopoietic hierarchy.

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Acknowledgements

We thank J. Yetz-Aldape for cell sorting; R. Czyzewski for mouse husbandry; X. Qi for technical support; and B.A. Morgan and the Georgopoulos laboratory for critical reading of the manuscript. Supported by the National Institutes of Health (5R37 R01 AI33062 to K.G. and 5T32 AI07529 to S.Y.N.), the Human Frontier Science Program (T.Y.) and the Transplantation Biology Research Center at Massachusetts General Hospital (S.Y.N.).

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Competing interests

The authors declare no competing financial interests.

Correspondence to Katia Georgopoulos.

Supplementary information

  1. Supplementary Fig. 1

    Distribution of the Ikaros-GFP reporter in the wild-type (WT) and Ikaros-null (Null) LSK and LK compartments. (PDF 69 kb)

  2. Supplementary Fig. 2

    Post-sort purity of the LK-GFPhi population. (PDF 655 kb)

  3. Supplementary Fig. 3

    Kinetics of T and B cell development from wild-type Ikaros-GFP LSK populations. (PDF 536 kb)

  4. Supplementary Fig. 4

    Analysis of lineage—Ikaros-GFP subsets for expression of CD34, and CD16/32 and Flt3 on various genetic backgrounds. (PDF 562 kb)

  5. Supplementary Fig. 5

    Cell fate decisions in the early hematopoietic hierarchy and their regulation by Ikaros. (PDF 334 kb)

  6. Supplementary Table 1

    Limiting dilution analysis of T cell differentiation from wild-type and Ikaros-null LSK cells. (PDF 20 kb)

  7. Supplementary Table 2

    Primers and PCR conditions used to examine lineage-specific genes expression in LK and LSK populations (PDF 83 kb)

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Figure 1: Molecular and functional subdivision of the bone marrow HSC compartment with the Ikaros reporter.
Figure 2: Molecular and functional subcategorization of erythro-myeloid progenitors by the Ikaros reporter.
Figure 3: Molecular and functional subcategorization of Ikaros-null LSK cells with the Ikaros reporter.
Figure 4: Clonal analysis of wild-type and Ikaros-null LMPPs.
Figure 5: Molecular and functional analysis of Ikaros-null erythro-myeloid progenitors.