Effects of WT and mutant C/EBPα on C/EBPα–/– cells. (A) NE mRNA transcripts in C/EBPα–/– cells ectopically expressing p42 or mutant Lys298Glu C/EBPα. Total RNA was extracted from GFP-sorted C/EBPα–/– cells expressing the indicated C/EBPα protein. NE transcripts were detected by RT-PCR (22, 25, and 28 cycles), and identity of the PCR products was confirmed by Southern blot hybridization using a 32P-labeled internal NE oligonucleotide probe. Panel 3 shows the ethidium bromide–stained agarose gel in which PCR products (28 cycles) were run. Amplification of GRB2 mRNA transcripts was performed as loading control. (B) Morphology (May-Grünwald/Giemsa–stained cytospins; original magnification, × 40.) of GFP-selected C/EBPα–/– cells transduced with the MigR1 or C/EBPα (wild-type or mutant) retrovirus and incubated with G-CSF for 4 days (lower panel), or left in their culture medium that contains 2% IL-3 and 2% SCF and is indicated as IL-3 (upper panel). Representative of 3 independent experiments.
