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STEM_232_sm_suppinfofigS1.tif36KSupplementary Figure 1. Clonogenic myeloid colony readout in methylcellulose of 100 plated CMPs (Lin(−/lo)cKit+Sca1-IL7Rα−CD34+FcγR(lo)) FACS-purified from wild-type (+/+) and mutant (Plstc/Plstc) E14.5 FLs.
STEM_232_sm_suppinfofigS2.tif35KSupplementary Figure 2. Clonogenic myeloid colony readout in methylcellulose of 100 plated Lin(−/lo)cKit+Sca1-IL7Rα−CD34-FcγR(hi) FACS-purified cells from wild-type (+/+) and mutant (Plstc/Plstc) E14.5 FLs.
STEM_232_sm_suppinfofigS3.tif393KSupplementary Figure 3. Phenotype of Plstc/Plstc embryo and wild-type sibling in yolk sac at E15.5 showing a marked deficit of red cells in the vitelline and umbilical circulation.
STEM_232_sm_suppinfofigS4.tif135KSupplementary Figure 4. Representative FACS plots of E14.5 fetal liver after 28 days culture on OP9-DL1 (A) and OP9 (B) stromal cell lines.
STEM_232_sm_suppinfofigS5.tif80KSupplementary Figure 5. Ikaros plays a crucial role in the self-renewal machinery for the ancestral pool of the long-term reconstituting subset of HSCs. E12.5 Plastic homozygotes boast normal numbers of KTLS(CD150+) LT-HSCs which disappear by E14.5-15.5 leaving accumulated numbers of all non-long-term self-renewing cellular subsets, including KTLS(CD150-) cells. Red arrows indicate the other crucial differentiation checkpoints where Ikaros plays an important role: (i) in the cell fate choice of MPPs/LMPPs to become either lymphoid or myeloid cells, Ikaros regulates the lymphoid checkpoint; and (ii) in the choice of CMPs to become GMPs or MEPs, Ikaros regulates the erythroid checkpoint. With several independent lines of evidence emerging that multipotent LT-HSCs may first lose megakaryocyte/erythrocyte potential and that there may be a more intimate and direct connection between these two cell subsets, Ikaros would clearly regulate this early developmental cell fate choice.
STEM_232_sm_suppinfotableS1.doc61KTable S1. Sequences of primers used to examine gene expression by qRT-PCR in FACS-purified cells in this study
STEM_232_sm_suppinfo.doc23KSupporting Information

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