Flow-cytometric phosphoprotein analysis reveals agonist and temporal differences in responses of murine hematopoietic stem/progenitor cells

PLoS One. 2008;3(11):e3776. doi: 10.1371/journal.pone.0003776. Epub 2008 Nov 20.


Hematopoietic stem cells (HSCs) are probably the best-studied adult tissue-restricted stem cells. Although methods for flow cytometric detection of phosphoproteins in hematopoeitic progenitors and mature cells are available, analogous protocols for HSC are lacking. We present a robust method to study intracellular signaling in immunophenotypically-defined murine HSC/progenitor cell (HPC)-enriched populations. Using this method, we uncover differences in the response dynamics of several phosphoproteins representative of the Ras/MAP-Kinase(K), PI3K, mTOR and Jak/STAT pathways in HSC/HPCs stimulated by Scf, Thpo, as well as several other important HSC/HPC agonists.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antigens, CD34 / biosynthesis
  • Cell Separation
  • Flow Cytometry / methods*
  • Hematopoietic Stem Cells / cytology*
  • Humans
  • Immunophenotyping
  • Kinetics
  • Ligands
  • Mice
  • Phosphoproteins / agonists
  • Phosphoproteins / chemistry*
  • Phosphorylation
  • Stem Cells / cytology*
  • Time Factors


  • Antigens, CD34
  • Ligands
  • Phosphoproteins