Induced expression of C/EBPε by AML1-MTG8.
(A) Northern blot analyses showing the expression of C-EBPε mRNA. Total RNAs (5 μg of RNA) were prepared from L-G infectants cultured with IL-3, blotted, and hybridized with 32P- labeled human C/EBPε cDNA, mouse PU.1 cDNA, and human G3PDH cDNA. (B) Northern blot analyses of poly(A)+RNAs from hematopoietic cell lines. Northern blots were hybridized with human C/EBPα cDNA and human C/EBPε cDNA. (C) Gel mobility shift assay showing the binding activities of cell extracts from L-G infectants and in vitro synthesized C/EBPε protein. (D) AML1-MTG8 activates the promoter of the C/EBPε gene. Cells were cotransfected with pGL3-C/EBPε (1280 bp) and either 0.2 μg of LNSX, 0.2 μg of LNSX-AML1b, or various concentrations of LNSX-AML1-MTG8. LNSX-AML1-MTG8 was added at 0.05 μg, 0.1 μg, and 0.2 μg in lanes 2, 3, and 4, respectively.
