Analysis of Bcl11a binding sites in mouse pro–B cells. (A) qPCR validation of Bcl11a binding at the Mdm2, Mdm4, and Bcl2 loci in ChIP assay. Potential Bcl11a binding sites (red boxes) at the Mdm2 and Mdm4 loci, predicted as GGCCGG-containing sequences. Black bars indicate the qPCR-amplified regions of the validated sites. Potential Bcl11a binding sites (arrows) at the mouse Bcl2 locus, predicted as homologous regions to BCL11A-binding sites at the human BCL2 locus (ENCODE project), are analyzed by ChIP assay. The fold-enrichments of the amplified regions are (from 5′ to 3′) are as follows: 1.31 ± 0.40, 1.69 ± 0.32, 5.32 ± 0.92 (red arrow), 1.87 ± 0.001, 2.37 ± 0.60, 2.80 ± 0.43, 1.32 ± 0.21, 2.12 ± 0.44, 1.88 ± 0.21, 2.29 ± 0.40, 3.00 ± 0.16, and 2.68 ± 0.72. (B) Distribution of Bcl11a-binding sites in the genome from ChIP-Seq analysis. The 10-kb region upstream from transcription start site is defined as Proximal Promoter, and the 10-kb region downstream from transcription stop site is defined as downstream. (C) qPCR validation of Bcl11a binding at the genomic loci of Bcl-xL, p21, Pax5, and E2A. Genomic DNA pull-down using IgG was used as a control. A–C represent three independent experiments. *, P < 0.05; **, P < 0.01.