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Blood, Vol. 103, Issue 10, 3615-3623, May 15, 2004
PU.1 determines the self-renewal capacity of erythroid progenitor cells
Blood Back et al.
103: 3615
Supplemental materials for: Back et al, Vol 103, Issue 10, 3615-3623
Files in this Data Supplement:
- Figure S1. The PU.1G mutation is null. (JPG, 40.8 KB)
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(A) Southern blot analysis of the PU.1 mutations. EcoRI-digested tail DNA was analyzed using probe B (see Fig. 1A). (B) Absence of PU.1 mRNA in 14.5 dpc PU.1G/G FL cells. PU.1 mRNA was detected by RT-PCR, using the indicated primers (see Fig. 1A). (+) or (–) RT indicates the presence or absence of reverse transcriptase in the reaction. C) Western blot analysis of PU.1 expression from nuclear extracts of 15.5 dpc FL cells. D) Lymphoid and myeloid populations in PU.1G/G FLs. 17.5 dpc FL cells were stained for the B lymphoid markers CD19 and CD43 (top), and for the myeloid markers CD11b and Gr1 (bottom). The percentage of each gated population is indicated. Results are representative of more than 3 experiments for this stage. Similar results were obtained from earlier stages (14.5-17.5 dpc). E) The GFP protein is a reliable reporter for PU.1 expression. Bone marrow myeloid, splenic B and thymic T cells from adult heterozygous PU.1+/G mice were stained with antibodies specific for the indicated markers (dot plots), and the gated populations were analyzed for GFP expression (histograms). The gray histograms indicate background fluorescence of the same populations from WT mice. Similar results were obtained from more than 10 experiments.
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