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Article first published online: 12 FEB 2013
Copyright © 2012 AlphaMed Press
Volume 31, Issue 2, pages 293–304, February 2013
How to Cite
Bourdeau, A., Trop, S., Doody, K. M., Dumont, D. J. and Tremblayef, M. L. (2013), Inhibition of T Cell Protein Tyrosine Phosphatase Enhances Interleukin-18-Dependent Hematopoietic Stem Cell Expansion. STEM CELLS, 31: 293–304. doi: 10.1002/stem.1276
Author contributions: A.B.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript; S.T. and K.M.D.: collection and/or assembly of data, data analysis and interpretation, manuscript writing; D.J.D.: data interpretation, manuscript writing; and M.L.T.: conception and design, financial support, data analysis and interpretation, manuscript writing, final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS November 8, 2012.
- Issue published online: 12 FEB 2013
- Article first published online: 12 FEB 2013
- Accepted manuscript online: 8 NOV 2012 06:54AM EST
- Manuscript Accepted: 11 OCT 2012
- Manuscript Received: 10 APR 2012
- Lymphoma Foundation
- Jeanne and Jean-Louis Lévesque Chair in Cancer Research
- Canadian Cancer Society
Vol. 31, Issue 6, 1224, Article first published online: 22 MAY 2013
Additional Supporting Information may be found in the online version of this article.
|sc-12-0345_sm_SupplTable1.pdf||8K||Supplementary Table 1|
|sc-12-0345_sm_SupplFigure1.tif||2946K||Supplemental Figure 1. Characterization of bone marrow hematopoietic stem cells. Further characterization of tc-ptp+/+ Lin−CD117+Sca-1+CD105+ and Lin−CD117+Sca-1+CD105- bone marrow stem cells subpopulations by flow cytometry showing expression of CD150, Flk-2 and CD34.|
|sc-12-0345_sm_SupplFigure2.tif||901K||Supplemental Figure 2. Assessment of apoptosis in hematopoietic stem cells. tc-ptp+/+ and tc-ptp−/− bone marrow (BM) were stained for hematopoietic stem cells as well as Annexin V and propidium iodide (PI) for analyzed by flow cytometry. The results shown are represented as Annexin V and PI and are gated on BM hematopoietic stem cells (HSC), which were identified as Lin-CD117+Sca-1+CD105+ cells.|
|sc-12-0345_sm_SupplFigure3.tif||858K||Supplemental Figure 3. Inhibition of tc-ptp by RNAi treatment. Mouse and human hematopoietic stem cells were electroporated without RNAi (PBS), with a control scramble sequence (SCR), or with TC-PTP RNAi (TC1 and/or TC2). RNA was isolated 72h post treatment from (A) Lin−CD117+Sca-1+CD105+ hematopoietic stem cells from the bone marrow of Balb/c mice and (B) Lin-CD34+CD133+ hematopoietic stem cells from human bone marrow. RT-PCR was performed to analyze levels of TC-PTP. GAPDH was used as a loading control.|
|sc-12-0345_sm_SupplFigure4.tif||1118K||Supplemental Figure 4. Further increased cytokine signaling in tc-ptp−/− Lin−CD117+Sca-1hi CD105+ subpopulation. Flow cytometry analysis of bone marrow Lin−CD117+Sca-1+ CD105+ cells from tc-ptp+/+ mice (thin line), tc-ptp−/− Lin−CD117+Sca-1+/hiCD105+ (thick line) and from tc-ptp−/− Lin−CD117+Sca-1hiCD105+ (extra-thick line) was performed to assess intracellular expression of (A) IL-18, (B) IL-18 binding protein (IL-18bp), or (C) phosphorylated Stat1 (p- Stat1) and total Stat1 protein. To better compare subpopulations, both the thin and thick line in all histograms have been reproduced as shown in Figure 5. A further overlay of the tc-ptp−/− Lin− CD117+Sca-1hiCD105+ (extra-thick line) population was performed. Nonspecific IgG was used as negative control (A, B, C: first flow cytometry histogram). The relative cell number, expressed as % max, is plotted against IgG, IL-18, IL-18bp, p-Stat1 or Stat1 fluorescence to compensate for the difference in cellularity between the populations. The mean fluorescence intensity (MFI) is indicated in the corresponding plot.|
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