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. 2004 Jul;24(13):5710–5720. doi: 10.1128/MCB.24.13.5710-5720.2004

FIG. 4.

FIG. 4.

Reprogramming of histone modifications in the promoter regions of several somatic tissue-specific genes. (A) Histone modifications at the Nfm promoter region (black bar). DNA prepared from chromatin immunoprecipitated with antibodies against AcH3, AcH4, H3-K4 dimethylation (H3-K4me2), and H3-K9 dimethylation (H3-K9me2) was PCR amplified with a primer set specific to the Nfm promoter region. (B) Real-time quantitative PCR analysis of Nfm. Histograms show the relative amount of immunoprecipitated DNA to input DNA for AcH3 and H3-K4me2. (C) Determination of the origin of immunoprecipitated DNA. The origin of PCR products was determined by sensitivity to MslI digestion. The thymocyte-derived products (mol) remain undigested (300 bp), whereas the ES cell-derived products (dom) are cut into two fragments (240 and 60 bp). (D) Histone modifications at the Nfl promoter region (black bar). DNA prepared from chromatin immunoprecipitated with antibodies to AcH3, AcH4, H3-K4me2, and H3-K9me2 was PCR amplified with a primer set specific to the Nfl promoter region. (E) Histone modifications at the Thy-1 promoter region (black bar). DNA prepared from chromatin immunoprecipitated with antibodies to AcH3, AcH4, H3-K4me2, and H3-K9me2 was PCR amplified with a primer set specific to the Thy-1 promoter region. (F) Real-time quantitative PCR analysis of Thy-1. Histograms show the relative amount of immunoprecipitated DNA to input DNA for AcH3, AcH4, and H3-K4me2. (G) Determination of the origin of immunoprecipitated DNA. DNA prepared from chromatin immunoprecipitated with antibodies to H3-K4me2 was PCR amplified with a primer set specific to the Thy-1 promoter region. The origin of PCR products was determined by sequencing of DNA extracted from independent clones. An adenine residue in the ES cell-derived genome (dom) is replaced with a thymine residue in the thymocyte-derived genome (mol). (H) Histone modifications at the c-myc promoter region (black bar). DNA prepared from chromatin immunoprecipitated with antibodies to AcH3, AcH4, H3-K4me2, and H3-K9me2 was PCR amplified with a primer set specific to the c-myc promoter region.