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Blood, Vol. 107, Issue 7, 2662-2672, April 1, 2006
Core erythropoietin receptor signals for late erythroblast development
Blood Menon et al.
107: 2662
Supplemental materials for: Menon et al
Files in this Data Supplement:
- Figure S1. Epo activation of Stat1 in wild-type-EpoR erythroblasts (JPG, 45.5 KB)
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Erythroblasts were expanded from wt-EpoR, EpoR-HM and EpoR-H bone marrow preparations. Isolated CD71highTer119neg cells then were washed, incubated for 6 hours in the absence of hematopoietic cytokines, and stimulated with Epo for 10 minutes. Lysates then were prepared directly, and were analyzed via western blotting for levels of activated PY-Stat1 and total Stat1. In wt-EpoR erythroblasts, Epo-activation of Stat1 was detected, but only upon prolonged ECL exposure.
- Figure S2. Epo induction of Cis-1 transcript expression in wt-EpoR and EpoR-H, but not EpoR-HM erythroblasts (JPG, 67.1 KB)
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Erythroblasts were prepared from wild-type EpoR, EpoR-H, and EpoR-HM bone marrow. Expanded cells were washed, incubated for 6 hours in the absence of hematopoietic cytokines and stimulated with Epo for the indicated intervals. Cells then were lysed in Trizol reagent and total RNA was isolated. Cis-1 transcript levels were analyzed by quantitative RT-PCR.
- Figure S3. p38 MAPK activation via minimal Epo receptor alleles (JPG, 68.2 KB)
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wt-Epo-R, EpoR-HM, and EpoR-H erythroblasts were expanded for 3 days. CD71highTer119neg cells were then isolated, washed, and incubated for 6 hours in the absence of hematopoietic cytokines. At the subsequent indicated intervals of Epo-exposure, lysates were prepared and analyzed by western blotting for levels of activated and total p38-MAPK.
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