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Blood, Vol. 108, Issue 3, 896-903, August 1, 2006
Proper levels of c-Myb are discretely defined at distinct steps of hematopoietic cell development
Blood Sakamoto et al.
108: 896
Supplemental materials for: Sakamoto et al, Vol 108, Issue 3, 896-903
Files in this Data Supplement:
- Table S1. Differential counts of hematopoietic cells derived from WT and c-mybTet/KO ES cells (PDF, 19.3 KB) -
VE-cadherin+ CD31+ cells were sorted from differentiating wild-type ES cells and c-mybTet/KO ES cells as described in Figure 2. Cells were recultured for 9 days with OP9 cells and cytokines (SCF, IL3, Epo) in the presence of different concentrations of Tet. Floating cells were harvested and stained with May-Grünwald Giemsa solution. M indicates monocytes/macrophages; PMN, polymorphonuclear leukocytes; Meg, megakaryocytes; EryB, basophilic erythroblasts; and EryC, enucleated erythrocytes.
- Figure S1. Expression of endogenous c-Myb in differentiating ES cells (JPG, 32.2 KB)
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Wild-type ES cells (WT) and c-myb−/− ES cells (KO) were allowed to differentiate on OP9 stromal cell layers for 4 days. Flk-1+ cells were sorted by FACS and recultured with OP9 cells for 3 days. Cells were then lysed and subjected to immunoprecipitation and Western blotting with an antibody against c-Myb. Two isoforms of c-Myb, p75 (closed triangle) and p89 (open triangle), were detected in WT cells. Cell lysate from OP9 stromal cell layers cultured without ES cells was also analyzed as a negative control (OP9).
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