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Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
sc-12-0345_sm_SupplData.pdf17KSupplementary Data
sc-12-0345_sm_SupplTable1.pdf8KSupplementary Table 1
sc-12-0345_sm_SupplFigure1.tif2946KSupplemental 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.tif901KSupplemental 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.tif858KSupplemental 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.tif1118KSupplemental 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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