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. 1998 Aug 17;188(4):699–713. doi: 10.1084/jem.188.4.699

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Increased expression of B lineage–associated genes in E12- expressing macrophages. (A) Northern blot analysis of EBF, Pax-5, IL7Rα, λ5, c-fms, Id2, and actin mRNA expression. 10 μg of total RNA extracted from the 70Z/3 pre-B, macrophage, m/neo, and the E12-expressing macrophage clones 2C1, 2C6, and D3 was electrophoresed through an 0.8% agarose gel and transferred to nylon membrane. The blots were probed sequentially with ³²P-labeled cDNA probes for the indicated genes. (B) RT-PCR analysis of Rag-1 and Ikaros RNA expression. 2 μg of total RNA was reverse transcribed using an oligo dT₁₅ primer in the presence (+) or absence (−) of RT. 100 ng of cDNA was used for PCR amplification with primers specific for Rag-1 (25 cycles), the COOH-terminal domain of Ikaros (25 cycles), or actin (13 cycles; C) μE5, EBF, Pax-5, and Oct DNA binding activity in E12-expressing macrophage cell lines. 10 μg of nuclear extract from each of the indicated cell lines was incubated with a ³²P-labeled oligonucleotide probe containing the binding site for E2A (μE5), EBF, Pax-5, and Oct.

Increased expression of B lineage–associated genes in E12- expressing macrophages. (A) Northern blot analysis of EBF, Pax-5, IL7Rα, λ5, c-fms, Id2, and actin mRNA expression. 10 μg of total RNA extracted from the 70Z/3 pre-B, macrophage, m/neo, and the E12-expressing macrophage clones 2C1, 2C6, and D3 was electrophoresed through an 0.8% agarose gel and transferred to nylon membrane. The blots were probed sequentially with ³²P-labeled cDNA probes for the indicated genes. (B) RT-PCR analysis of Rag-1 and Ikaros RNA expression. 2 μg of total RNA was reverse transcribed using an oligo dT₁₅ primer in the presence (+) or absence (−) of RT. 100 ng of cDNA was used for PCR amplification with primers specific for Rag-1 (25 cycles), the COOH-terminal domain of Ikaros (25 cycles), or actin (13 cycles; C) μE5, EBF, Pax-5, and Oct DNA binding activity in E12-expressing macrophage cell lines. 10 μg of nuclear extract from each of the indicated cell lines was incubated with a ³²P-labeled oligonucleotide probe containing the binding site for E2A (μE5), EBF, Pax-5, and Oct.