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Blood, Vol. 108, Issue 5, 1524-1532, September 1, 2006
Lyn kinase promotes erythroblast expansion and late-stage development
Blood Karur et al.
108: 1524
Supplemental materials for: Karur et al
Files in this Data Supplement:
- Figure S1. Bone marrow derived Lyn-/- erythroid progenitor cells falter in their ex vivo expansion and development to KitnegCD71high erythroblasts (JPG, 287 KB)
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A. Kitpos progenitors were prepared from bone marrow of Lyn-/- and Lyn+/+ mice, and expanded in SP34-EX medium as detailed in Methods (and the legend to Figure 3). At the indicated time-points, the formation of Kit (CD117) - positive and/or CD71- positive erythroblasts was assayed. Results shown are for two independent experiments (#2, #3). B. Expansion of erythroid progenitor cells also was monitored by direct cell counts. C. Kitpos and Kitneg erythroblast formation frequencies also are illustrated in ratio plots.
- Figure S2. Lyn-deficient erythroid progenitor cells exhibit attenuated rates of Epo and SCF dependent 3HdT- incorporation (JPG, 89.2 KB)
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A. For Lyn+/+ and Lyn-/- mice, KitposCD71high progenitor cells were isolated from bone marrow expansion cultures. Cells then were cultured in SP34-EX medium for 24 hours in the indicated cytokines, and pulsed with 3HdT. Incorporation rates shown are for n=4 Lyn+/+ and Lyn-/- mice. B. Also graphed are response to combined doses of Epo and SCF which supported more-than-additive responses.
- Figure S3. SCF and Epo activation of ERKs is unaffected due to Lyn deficiency (JPG, 74.3 KB)
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KitposCD71high and KitnegCD71high erythroblasts were expanded from Lyn+/+ and Lyn-/- mice. Cells were then deprived of cytokines for six hours, and were exposed to Epo plus SCF (5U/mL; 100ng/mL). At the indicated intervals, levels of phospho-ERKs (and total ERKs) were assayed by western blotting.
- Figure S4. In bone marrow-derived erythroid progenitor cells, Lyn deficiency is associated with enhanced Akt activation (JPG, 108 KB)
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KitnegCD71high and KitposCD71high erythroblasts were expanded from Lyn+/+ and Lyn-/- mice. Cells were then deprived of cytokines for six hours, and were exposed to Epo plus SCF (5U/mL; 100ng/mL). At the indicated intervals, levels of phospho-Akt (and total Akt) were assayed by western blotting.
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