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. Author manuscript; available in PMC: 2013 Jul 1.
Published in final edited form as: Stem Cells. 2012 Jul;30(7):1447–1454. doi: 10.1002/stem.1127

Figure 2. HSCs are highly responsive to SCF and TPO stimulation.

Figure 2

(A) Flow chart of the developed “HSC phospho-flow” method. (B) Sca1+ enriched, CD150+ CD41− bone marrow cells were serum- and cytokine-starved for 30 minutes and stimulated with 10 ng /ml of SCF or 20 ng/ml of TPO for 10 minutes at 37°C. Levels of phosphorylated ERK1/2, Akt, Stat3, and Stat5 were measured using phospho-specific flow cytometry. HSCs (defined as [Lin CD48]−/low cKit+ cells) were gated for data analysis. Representative gating strategy and plots of phosphorylated signaling proteins are shown (n= 6).