Figure 5
Figure 5. Coexpression of both C/EBPα-Cm and C/EBPα-Nm led to AML with leukocytosis with shorter latencies. (A) Kaplan-Meier analysis for the survival of mice that received transplants of BM cells transduced with both Myc-tagged C/EBPα-Cm-IG and pMYs-IR (Myc-Cm/pMYs-IR, n = 6), both pMYs-IG and Flag-tagged C/EBPα-Nm-IR (pMYs-IG/Flag-Nm, n = 6), both Myc-tagged C/EBPα-Cm-IG and Flag-tagged C/EBPα-Nm-IR (Myc-Cm/Flag-Nm, n = 11), or mock (pMYs-IG/pMYs-IR, n = 8). (B) Cytospin preparations of BM cells derived from mice/My-Cm/pMYs-IR or mice/Myc-Cm/Flag-Nm were stained with Giemsa. A representative photograph is shown. Images were obtained with a BX51 microscope and a DP12 camera (Olympus); objective lens, UplanFl (Olympus); original magnification ×100. (C) Flow cytometric analysis of BM cells derived from mice/Myc-Cm/pMYs-IR (left) or mice/Myc-Cm/Flag-Nm (right). The dot plots show expression of dsRED versus expression of GFP (1st panel). In the indicated gating, the dot plots show expression of Gr-1, CD11b, B220, CD19, or c-kit labeled with phycoerythrin-Cy5–conjugated streptavidin versus expression of GFP. (D) Expression of Myc-tagged C/EBPα-Cm protein and p30 protein generated by Flag-tagged C/EBPα-Nm in BM cells derived from mice/pMYs-IG/pMYs-IR (lane 1), mice/Myc-Cm/pMYs-IR (lanes 2-3), or mice/Myc-Cm/Flag-Nm (lanes 4-5) was detected by using anti-Myc monoclonal Ab (top) and ant-Flag mAb (middle), respectively, in Western blot analysis. Equal loading was evaluated by probing the immunoblots with anti-tubulin Ab (bottom). Data are representative of 3 independent experiments. (E) Peripheral blood smears obtained from mice/Myc-Cm/pMYs-IR (left) or mice/Myc-Cm/Flag-Nm (right) were stained with Giemsa. Images were obtained with a BX51 microscope and a DP12 camera (Olympus); objective lens, UplanFl (Olympus); original magnification ×20. (F) Counts of white blood cells (WBC) obtained from mice/Myc-Cm/pMYs-IR (n = 6), mice/Myc-Cm/Flag-Nm (n = 8), or mice/pMYs-IG/pMYs-IR (n = 8). All data points correspond to the mean and the standard deviation (SD). Statistically significant differences are shown. *P < .05.

Coexpression of both C/EBPα-Cm and C/EBPα-Nm led to AML with leukocytosis with shorter latencies. (A) Kaplan-Meier analysis for the survival of mice that received transplants of BM cells transduced with both Myc-tagged C/EBPα-Cm-IG and pMYs-IR (Myc-Cm/pMYs-IR, n = 6), both pMYs-IG and Flag-tagged C/EBPα-Nm-IR (pMYs-IG/Flag-Nm, n = 6), both Myc-tagged C/EBPα-Cm-IG and Flag-tagged C/EBPα-Nm-IR (Myc-Cm/Flag-Nm, n = 11), or mock (pMYs-IG/pMYs-IR, n = 8). (B) Cytospin preparations of BM cells derived from mice/My-Cm/pMYs-IR or mice/Myc-Cm/Flag-Nm were stained with Giemsa. A representative photograph is shown. Images were obtained with a BX51 microscope and a DP12 camera (Olympus); objective lens, UplanFl (Olympus); original magnification ×100. (C) Flow cytometric analysis of BM cells derived from mice/Myc-Cm/pMYs-IR (left) or mice/Myc-Cm/Flag-Nm (right). The dot plots show expression of dsRED versus expression of GFP (1st panel). In the indicated gating, the dot plots show expression of Gr-1, CD11b, B220, CD19, or c-kit labeled with phycoerythrin-Cy5–conjugated streptavidin versus expression of GFP. (D) Expression of Myc-tagged C/EBPα-Cm protein and p30 protein generated by Flag-tagged C/EBPα-Nm in BM cells derived from mice/pMYs-IG/pMYs-IR (lane 1), mice/Myc-Cm/pMYs-IR (lanes 2-3), or mice/Myc-Cm/Flag-Nm (lanes 4-5) was detected by using anti-Myc monoclonal Ab (top) and ant-Flag mAb (middle), respectively, in Western blot analysis. Equal loading was evaluated by probing the immunoblots with anti-tubulin Ab (bottom). Data are representative of 3 independent experiments. (E) Peripheral blood smears obtained from mice/Myc-Cm/pMYs-IR (left) or mice/Myc-Cm/Flag-Nm (right) were stained with Giemsa. Images were obtained with a BX51 microscope and a DP12 camera (Olympus); objective lens, UplanFl (Olympus); original magnification ×20. (F) Counts of white blood cells (WBC) obtained from mice/Myc-Cm/pMYs-IR (n = 6), mice/Myc-Cm/Flag-Nm (n = 8), or mice/pMYs-IG/pMYs-IR (n = 8). All data points correspond to the mean and the standard deviation (SD). Statistically significant differences are shown. *P < .05.

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