Figure 3.

Northern blot analysis of endogenous PEBP2αB1 and ear-2 mRNA in 32Dcl3 cells during granulocytic differentiation. Poly(A)⁺ RNAs were prepared from 32Dcl3 stably transfected with the pBabeneo vector alone at days 0, 3, 6, 8, and 9 after the addition of G-CSF. Five micrograms of poly(A)⁺ RNA was analyzed by Northern blotting using ³²P-labeled ear-2 cDNA as a probe. To detect an early differentiation maker, MPO, its cDNA was used as a probe for the same blot after it was stripped. A different membrane containing 2.5 μg of poly(A)⁺ RNA was hybridized with ³²P-labeled PEBP2αB1 cDNA as a probe. As an internal control, β-actin mRNA expression was analyzed for each membrane after striping. Positions of 18S and 28S rRNAs are indicated.