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1.
Figure 2

Figure 2. From: Histone hyperacetylation within the β-globin locus is context-dependent and precedes high-level gene expression.

Histone modifications across the β-globin locus during ES cell differentiation. The murine β-globin locus and locations of amplimers used for quantitative PCR are shown at the top, as in . (A) Enrichments for H4Ac and H3K4 dimethyl within the β-globin locus in undifferentiated ES cells. (B) Globin mRNA expression during days 2 to 8 of EB differentiation, normalized to 18s mRNA. (C) Benzidine staining during days 2 to 8 of EB differentiation. (D) Enrichments for H4Ac (top) and H3K4 dimethyl (bottom) in EBs differentiated for 2, 4, 6, and 8 days.

George Fromm, et al. Blood. 2009 October 15;114(16):3479-3488.
2.
Figure 7

Figure 7. From: Histone hyperacetylation within the β-globin locus is context-dependent and precedes high-level gene expression.

Domain formation after inhibition of RNA Pol II elongation by DRB. (A) DRB effect on mature and primary mRNA transcripts of the embryonic βh1 globin gene. (B) ChIP results for RNA Pol II at the promoters and the 3′ region of ϵy and βh1 in untreated (■) and DRB-treated () samples. (C) H3Ac (top) and H3K4 dimethyl (bottom) ChIP results for E12.5 peripheral blood across the region harboring the embryonic β-globin genes in untreated (□) and DRB-treated (■) cells.

George Fromm, et al. Blood. 2009 October 15;114(16):3479-3488.
3.
Figure 3

Figure 3. From: Histone hyperacetylation within the β-globin locus is context-dependent and precedes high-level gene expression.

Histone modifications within the β-globin locus in the absence of the LCR. ChIP assays were performed using anti-H3Ac and anti-H3K4 dimethyl antibodies in primary cells isolated from wild-type (WT) mice and mice homozygous for deletion of the endogenous β-globin LCR (ΔLCR). The β-globin locus and positions of PCR amplimers are shown to scale at the top. Enrichments in E12.5 embryonic blood for (A) H3Ac and (B) H3K4 dimethyl and in E14.5 fetal liver for (C) H3Ac and (D) H3K4 dimethyl are shown in bar graph format beneath. The more lightly shaded portion of the graph represents the region in which enrichments for the indicated histone modifications were significantly lower than in erythroid cells from wild-type mice.

George Fromm, et al. Blood. 2009 October 15;114(16):3479-3488.
4.
Figure 5

Figure 5. From: Histone hyperacetylation within the β-globin locus is context-dependent and precedes high-level gene expression.

Histone modifications within the β-globin locus in definitive erythroid cells from mice harboring deletions of the ϵy- and/or βh1-globin gene promoters. (A) Enrichments for H3Ac (top) and H3K4 dimethyl (bottom) at the indicated locations within the β-globin locus in E14.5 fetal liver from wild-type and Δϵy/Δβh1 Pr. mice. (B) Enrichments for H3Ac (top) and H3K4 dimethyl (bottom) at the indicated locations within the β-globin locus in E14.5 fetal liver from Δϵy Pr. and Δβh1 Pr. mice. The locus is shown to scale above the bar graphs as before, with the promoter deletions indicated by gray brackets. The more lightly shaded portions of the graphs in panels A and B represent regions in which a significant difference in histone modification levels between erythroid cells derived from wild-type and the promoter deletion mice is evident.

George Fromm, et al. Blood. 2009 October 15;114(16):3479-3488.
5.
Figure 4

Figure 4. From: Histone hyperacetylation within the β-globin locus is context-dependent and precedes high-level gene expression.

Histone modifications within the β-globin locus in primitive erythroblasts from mice harboring deletions of the ϵy- and/or βh1-globin gene promoters. Histone modification patterns and mRNA expression levels were analyzed in E12.5 peripheral blood from mice homozygous for the deletion of either one (Δϵy Pr. and Δβh1 Pr.) or both (Δϵy/Δβh1 Pr.) of the embryonic globin gene promoters. (A) Schematic of the β-globin locus showing a closer view of the regions being analyzed and the location of each deletion. (B) Globin expression levels using quantitative RT-PCR. Values are standardized to 18S mRNA levels and are presented as the fold change with respect to the levels seen in wild-type mice. (C) Enrichments for H3Ac (top) and H3K4 dimethyl (bottom) at the indicated locations within the β-globin locus in E12.5 blood from the indicated embryonic promoter deletion mice.

George Fromm, et al. Blood. 2009 October 15;114(16):3479-3488.
6.
Figure 6

Figure 6. From: Histone hyperacetylation within the β-globin locus is context-dependent and precedes high-level gene expression.

Histone modification state of a unique sequence with a different positioning within 2 alleles of the β-globin locus. (A) Schematic of the region of the β-globin locus between the β1- and β2-globin genes for the D and S alleles. The block of unique sequence that occupies different relative positions within each allele is indicated by the thicker bar with a light fill, whereas the large insertions of repetitive DNA specific for each allele are indicated by thicker bars with dark fill. Positions of PCR amplimers used in analysis of ChIP assays are shown by the short bars beneath the representation of each allele. * represents the amplimer located within the unique sequence; “X,” an amplimer within a unique region that is found in the D but not the S allele. (B) Bar graphs showing enrichment levels for H3Ac (top) and H3K4 dimethyl (bottom), relative to inactive gene loci. Bars within the dark-shaded regions represent portions of the locus that are common to both the D and S alleles; lightly shaded regions, portions of the locus unique to the D or S alleles. * represents enrichments observed for the translocated unique sequence common to D and S; “X,” enrichment observed for the sequence unique to D.

George Fromm, et al. Blood. 2009 October 15;114(16):3479-3488.
7.
Figure 1

Figure 1. From: Histone hyperacetylation within the β-globin locus is context-dependent and precedes high-level gene expression.

Histone modifications across the β-globin locus in MEL745a and FDCP-Mix cells. The murine β-globin locus is depicted to scale as a line at the top, with the 4 developmentally regulated β-globin genes labeled over black arrows, and exons represented by black bars on the line. Unlabeled, thicker black bars on the line represent β-globin pseudogenes. HS2 indicates hypersensitive site 2 of the locus control region (LCR). Black bars below the line depict the locations of PCR amplimers used in the ChIP assay, which is shown in turn in bar graph format for each of the antibodies used for IP. (A) Quantitative RT-PCR of combined β1- and β2-globin mRNA expression levels relative to 18s mRNA in FDCP-Mix, and in both uninduced (Un-Ind.) MEL745a and MEL745a induced with 2% DMSO for 72 hours (Ind.). (B) Fold induction of β1- and β2-globin mRNA expression levels in MEL745a on culture in 2% DMSO for 72 hours, relative to uninduced MEL745a. (C) Bar graph depicting H3Ac and (D) H3K4 dimethyl enrichment levels, relative to inactive gene loci (“ChIP”), in both uninduced (Un-ind.) and induced (Ind.) MEL745a. The positions of each bar correspond to the positions of the corresponding amplimers shown to scale at the top. (E) H3K4 trimethyl enrichment levels, relative to inactive gene loci, for uninduced and induced MEL745a. (F) H3Ac and H3K4 dimethyl enrichment levels for FDCP-Mix.

George Fromm, et al. Blood. 2009 October 15;114(16):3479-3488.

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