Figure 6
Dmtf1 has both Arf-dependent and Arf-independent pathways. (A) Human Dmtf1-FL or Dmtf1-DN were retrovirally transduced into Jurkat T cells (Arf-null) and purified by GFP sorting. Cell-cycle analysis was measured with Hst staining. Results are shown as average ± SD (n = 3). *P < .05. (B) Cell numbers of Jurkat cells expressing FL or DN were determined daily. *P < .01. Results shown are from 1 of 3 independent experiments. (C) Retroviral vector MIG-Arf and MIG-Mock were transduced into WT or Dmtf1-KO Lin− cells, and GFP+ cells were sorted flow cytometrically and used in a CFU assay. Results are from 1 of 3 independent experiments. *P < .01 and **P < .03. (D) Analysis of gene expression in Dmtf1-KO KSLs by quantitative RT-PCR analysis of RNA isolated from sorted KSL cells from primary Dmtf1-KO or WT. The relative expression is shown normalized to gene expression in control HSCs (mean ± SD, n = 3). *P < .05. (E) Relative expressions of JunB in B and CD4+ T cells. Sorted splenic B cells and CD4+ T cells were analyzed by quantitative RT-PCR. The relative expression is shown normalized to gene expression in WT CD4 or B cells, respectively (mean ± SD, n = 3). *P < .05.

Dmtf1 has both Arf-dependent and Arf-independent pathways. (A) Human Dmtf1-FL or Dmtf1-DN were retrovirally transduced into Jurkat T cells (Arf-null) and purified by GFP sorting. Cell-cycle analysis was measured with Hst staining. Results are shown as average ± SD (n = 3). *P < .05. (B) Cell numbers of Jurkat cells expressing FL or DN were determined daily. *P < .01. Results shown are from 1 of 3 independent experiments. (C) Retroviral vector MIG-Arf and MIG-Mock were transduced into WT or Dmtf1-KO Lin cells, and GFP+ cells were sorted flow cytometrically and used in a CFU assay. Results are from 1 of 3 independent experiments. *P < .01 and **P < .03. (D) Analysis of gene expression in Dmtf1-KO KSLs by quantitative RT-PCR analysis of RNA isolated from sorted KSL cells from primary Dmtf1-KO or WT. The relative expression is shown normalized to gene expression in control HSCs (mean ± SD, n = 3). *P < .05. (E) Relative expressions of JunB in B and CD4+ T cells. Sorted splenic B cells and CD4+ T cells were analyzed by quantitative RT-PCR. The relative expression is shown normalized to gene expression in WT CD4 or B cells, respectively (mean ± SD, n = 3). *P < .05.

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