Figure 6
Figure 6. Mitotic quiescence of the primitive hematopoietic cells in the leukemic hosts and expression of several cell-cycle regulators. As illustrated in Figure 1A, 107 BMNCs from B6.SJL mice were transplanted with or without 106 Notch1-induced leukemia cells (CD45.2+GFP+) into lethally irradiated C57BL/6J recipients. At the 2-week time point, LKS cells from CD45.1+ BMNCS were sorted for staining with PI to assess the general cell-cycle profile (G0/G1 vs S/G2 + M) or staining with PY in conjunction with Hoechst 33342 (HO) to specifically determine the portion of cells in G0 vs G1 with flow cytometry. An aliquot of the cells was also used to examine the expression of several cell-cycle regulators with real time RT-PCR. (A) PI staining. A representative figure is shown from 1 of 2 experiments with similar results. (B) PY staining. Cells residing in G0 appear at the bottom of the G0/G1 peak as shown in the representative plot. Values are mean ± SD. **P = .017 (t test). (C) Expression of cell-cycle regulators. The CD45.1+ LKS cells were sorted directly into lysis buffer for real-time RT-PCR analysis. Data shown are the ratios between leukemic and control groups from 1 of 2 independent experiments with similar results. Values are mean ± SD. *P < .05 (t test). **P < .01 (t test).

Mitotic quiescence of the primitive hematopoietic cells in the leukemic hosts and expression of several cell-cycle regulators. As illustrated in Figure 1A, 107 BMNCs from B6.SJL mice were transplanted with or without 106Notch1-induced leukemia cells (CD45.2+GFP+) into lethally irradiated C57BL/6J recipients. At the 2-week time point, LKS cells from CD45.1+ BMNCS were sorted for staining with PI to assess the general cell-cycle profile (G0/G1 vs S/G2 + M) or staining with PY in conjunction with Hoechst 33342 (HO) to specifically determine the portion of cells in G0 vs G1 with flow cytometry. An aliquot of the cells was also used to examine the expression of several cell-cycle regulators with real time RT-PCR. (A) PI staining. A representative figure is shown from 1 of 2 experiments with similar results. (B) PY staining. Cells residing in G0 appear at the bottom of the G0/G1 peak as shown in the representative plot. Values are mean ± SD. **P = .017 (t test). (C) Expression of cell-cycle regulators. The CD45.1+ LKS cells were sorted directly into lysis buffer for real-time RT-PCR analysis. Data shown are the ratios between leukemic and control groups from 1 of 2 independent experiments with similar results. Values are mean ± SD. *P < .05 (t test). **P < .01 (t test).

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