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. 2014 Apr 7;211(4):635–642. doi: 10.1084/jem.20132398

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© 2014 Male et al.

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Figure 5. — E4bp4 protein directly binds the Eomes and Id2 loci. (A) Lineage-depleted bone marrow cells from either E4bp4^+/+ or E4bp4^−/− mice were cultured for 48 h in lymphocyte-inducing conditions, and Eomes, Id2, and T-bet transcripts were measured (six mice per genotype). *, P < 0.05. (B) Eomes and Id2 loci showing the location of the predicted E4bp4-binding sites. (C) ChIP analysis of the enrichment of E4bp4 at the indicated loci in chromatin from MNK-1 cells transduced with 5′ FLAG–E4bp4. C18 and E16 are polyclonal antibodies to E4bp4, and M2 is a monoclonal antibody to FLAG. GD (gene desert) was used as a negative control; Per2B was a positive control. (D) MNK-1 cells were transduced with either vector control or FLAG-E4bp4, and expression of Eomes and Id2 transcripts was determined by real-time PCR. (C and D) Data are representative of three independent biological replicates. Error bars show SEM.