Flow-cytometric phosphoprotein analysis reveals agonist and temporal differences in responses of murine hematopoietic stem/progenitor cells.
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.
Kalaitzidis
Demetrios
D
Hematology/Oncology-Cancer Biology Program, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA. dkalaitzidis@partners.org
Neel
Benjamin G
BG
D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis; D002469:Cell Separation; D005434:Flow Cytometry / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D016130:Immunophenotyping; D007700:Kinetics; D008024:Ligands; D051379:Mice; D010750:Phosphoproteins / Q000819:agonists / Q000737:chemistry*; D010766:Phosphorylation; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors
D018952:Antigens, CD34; D008024:Ligands; D010750:Phosphoproteins