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Blood, Vol. 110, Issue 12, 4101-4107, December 1, 2007
Histone acetylation at the human ß-globin locus changes with developmental age
Blood Yin et al.
110: 4101
Supplemental materials for: Yin et al
Files in this Data Supplement:
- Table S1. List of primers (PDF, 118 KB)
- Figure S1. Nucleosome histone binding measured by ChIP assay (JPG, 32.5 KB)
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Adult erythroid cells derived from CD34+ cells in two phase culture system were subject to ChIP analysis as described in “Materials and methods.” The specific antibodies recognize both acetylated and non-acetylated forms of histones H2A, H2B, H3, or H4. Four regions in the human beta-globin locus were estimated: HS2 (hHS2), epsilon gene promoter (hEpro), gamma gene promoter (hGpro), and beta gene promoter (hBpro). The amounts of immuno-precipitated DNA were corrected by the recovery of the external control gene mAire in corresponding ChIP assays.
- Figure S2. Measurement of non-specific immunoprecipitation in ChIP assay (JPG, 39.5 KB)
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Three types of control experiments were performed to evaluate non-specific immunoprecipitation in ChIP assay, i.e., mock experiments omitting chromatin, or antibody, or using normal IgG instead of the specific antibody. The figure shows the original data from one set of ChIP experiments using d100 fetal liver. The y-axis represents DNA amounts (ng) measured by real-time PCR. The x-axis indicates the estimated regions across the human  -globin locus. The mouse amylase control is shown in the most left data group. The filled bars: specific immunoprecipitation by the antibody against acetylated histone H3; striped bars: normal IgG control; grey bars: no antibody control; open bars: no chromatin control.
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