Protein expression and growth characteristics of 32Dcl3 cells expressing C/EBPε.
(A) Parental 32Dcl3 cells were washed and transferred from medium containing IL-3 to G-CSF, and cell lysates were prepared at the times indicated. Lysates of 32D/ε cells were obtained from cells growing in IL-3. Each protein extract (50 μg) was resolved on 4% to 20% gradient SDS-polyacrylamide gel and transferred to a nitrocellulose membrane. The membrane was probed with polyclonal antibody against C/EBPε. (B) Parental 32Dcl3 cells (solid symbols) and 32D/ε cells (open symbols) maintained in IL-3 were washed with PBS and transferred to medium containing 25 U/mL IL-3 (circle), 50 ng/mL G-CSF (square), or no cytokines (triangle). Viable cells were counted using the trypan blue dye exclusion method. Cells were diluted with each medium to keep cell density within 2 to 10 × 105/mL.
