American Society of Hematology

Figure 4

$Figure 4. Cells with impaired function of HPRT escape G2/M checkpoint arrest and apoptosis induced by 6TG. Fl5.12 cells transduced with Ctrl or 491 were cultured in the presence of 10 nM 6TG or no drug (UT). (A) At the indicated time points, a fraction of the culture was fixed, stained with PI, and analyzed by flow cytometry. The percentage of cells in the G2/M phase of the cell cycle (as defined by 4N DNA content) is graphed. **P < .01. ***P < .001. (B) Whole cell lysates from the indicated time points were subject to Western blot analysis with antibodies against p53 phosphorylated at serine 15 (P-p53; ab 9284; Cell Signaling Technology, Danvers, MA), HPRT, and actin. (C) After 3 days of treatment, a fraction of the culture was stained with fluorescein isothiocyanate-linked annexin V and PI and analyzed by flow cytometry. The percentages of apoptotic (annexin V+-PI−) and dead (annexin V+-PI+) cells are graphed. *P < .05. **P < .01.$

Cells with impaired function of HPRT escape G₂/M checkpoint arrest and apoptosis induced by 6TG. Fl5.12 cells transduced with Ctrl or 491 were cultured in the presence of 10 nM 6TG or no drug (UT). (A) At the indicated time points, a fraction of the culture was fixed, stained with PI, and analyzed by flow cytometry. The percentage of cells in the G₂/M phase of the cell cycle (as defined by 4N DNA content) is graphed. **P < .01. ***P < .001. (B) Whole cell lysates from the indicated time points were subject to Western blot analysis with antibodies against p53 phosphorylated at serine 15 (P-p53; ab 9284; Cell Signaling Technology, Danvers, MA), HPRT, and actin. (C) After 3 days of treatment, a fraction of the culture was stained with fluorescein isothiocyanate-linked annexin V and PI and analyzed by flow cytometry. The percentages of apoptotic (annexin V⁺-PI⁻) and dead (annexin V⁺-PI⁺) cells are graphed. *P < .05. **P < .01.

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