Figure 7
Figure 7. Ectopic Bcl-2 expression does not enhance colony formation of c-Mybf/d p210BCR/ABL-transduced HSCs. (A) Expression of c-Myb–regulated genes in p210BCR/ABL-transduced c-Mybf/f and c-Mybf/d progenitors. Western blot analysis of c-Myb, c-Myc, cyclin B1, and Bcl-2 expression in nontransduced and p210BCR/ABL-transduced enriched Lin− cells from c-Mybf/f and c-Mybf/d mice. Actin expression served as a control. (B) Colony formation of Bcl-2–transduced p210BCR/ABL-expressing c-Mybf/d progenitors. c-Mybf/d Lin−Sca-1+Kit+ cells were transduced with the MigRI/p210BCR/ABL retrovirus and 24 hours after retroviral transduction sorted for GFP expression. p210BCR/ABL-expressing c-Mybf/d Lin−Sca-1+Kit+ cells were subsequently transduced with a Bcl-2/MSCV/puro retrovirus or the empty MSCV/puro vector as a negative control. After 24 hours, cells (4 × 104/plate) were plated in methylcellulose in the presence of puromycin (1 μg/mL). Colony formation was assessed 9 days later. Representative of 3 different experiments.

Ectopic Bcl-2 expression does not enhance colony formation of c-Mybf/d p210BCR/ABL-transduced HSCs. (A) Expression of c-Myb–regulated genes in p210BCR/ABL-transduced c-Mybf/f and c-Mybf/d progenitors. Western blot analysis of c-Myb, c-Myc, cyclin B1, and Bcl-2 expression in nontransduced and p210BCR/ABL-transduced enriched Lin cells from c-Mybf/f and c-Mybf/d mice. Actin expression served as a control. (B) Colony formation of Bcl-2–transduced p210BCR/ABL-expressing c-Mybf/d progenitors. c-Mybf/d LinSca-1+Kit+ cells were transduced with the MigRI/p210BCR/ABL retrovirus and 24 hours after retroviral transduction sorted for GFP expression. p210BCR/ABL-expressing c-Mybf/d LinSca-1+Kit+ cells were subsequently transduced with a Bcl-2/MSCV/puro retrovirus or the empty MSCV/puro vector as a negative control. After 24 hours, cells (4 × 104/plate) were plated in methylcellulose in the presence of puromycin (1 μg/mL). Colony formation was assessed 9 days later. Representative of 3 different experiments.

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