Effect of Bcr-Abl on C/EBPα and G-CSFR mRNA expression.

Bcr-Abl reversibly blocks induction of C/EBPα and G-CSFR mRNA expression. (A-B) Expression of C/EBPα, G-CSFR, and PU.1 mRNA in 32Dcl3 and 32D_Bcr-Ablwt cells prior to and at day 3 after G-CSF stimulation as assessed by real-time PCR and semiquantitative RT-PCR. Real-time PCR (results shown in graphs) was performed as described in “Materials and methods.” The mRNA expression of C/EBPα, G-CSFR, and PU.1 mRNA was normalized to levels of G6PD mRNA. Induction of mRNA expression is shown as fold induction relative to unstimulated 32Dcl3 cells. Data from 1 of 2 independent experiments are shown. For semiquantitative RT-PCR (results shown in blots), real-time PCR reactions were stopped at appropriate time points (C/EBPα, 24 cycles; G-CSFR, 26 cycles; PU.1, 24 cycles; G6PD, 24 cycles). Aliquots were loaded onto agarose gels for further analyses. G6PD is shown as a control.