ITD mutations decrease C/EBPα expression.

(A) Real time RT-PCR analysis of 32D/Flt3-WT and 32D/Flt3-ITD stimulated with FL (●) or IL-3 (○) for the indicated times (hours). The relative expression was calculated in the same way as described for Figure 6. (B) Western blot analysis for C/EBPα from 32D cells expressing wild-type or ITD isoforms of Flt3. All cells were cytokine and serum starved for 12 hours prior to preparing the lysates for Western blot analyses. (C) Real time RT-PCR analysis for C/EBPα of FL-stimulated 32D/Flt3-WT and 32D/Flt3-ITD treated with the Flt3 kinase inhibitor D-65476. Cells were exposed to the indicated concentration of inhibitor for 12 hours. Mean and standard deviation of 2 independent experiments are shown.