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Blood, Vol. 113, Issue 7, 1547-1554, February 12, 2009
An insulator with barrier-element activity promotes  -spectrin gene expression in erythroid cells
Blood Gallagher et al.
113: 1547
Supplemental materials for: Gallagher et al
Files in this Data Supplement:
- Table S1. Oligonucleotide primers (PDF, 634 KB)
- Table S2. Electrophoretic mobility shift assay oligonucleotide probes (PDF, 623 KB)
- Table S3. Expression of α-spectrin/Aγ-globin mRNA in transgenic mice (PDF, 1.03 MB)
- Table S4. Developmental expression of Aγ-globin mRNA in AspEx1IVS1/Aγ-globin transgenic mice (PDF, 160 KB)
- Table S5. Patterns of Aγ-globin mRNA expression in tissues of AspEx1IVS1/Aγ-globin transgenic mice (PDF, 573 KB)
- Figure S1. Barrier element/Position effect variegation analyses in K562 cells (JPG, 40.8 KB)
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Top. Representative fluorescence-activated cell sorting histograms of individual clones are shown. (A) A clone with the HS2-β-globin promoter–GFP transgene without flanking sequences. (B) A clone with the HS2-β-globin promoter-GFP transgene flanked by α-spectrin intron 1′ sequences. (C) A clone with a single copy of the HS2-β-globin promoter-GFP transgene flanked by α-spectrin exon 1′ sequences. (D) A clone with two copies of the HS2-β-globin promoter-GFP transgene flanked by α-spectrin exon 1′ sequences. Bottom. Southern Blot analysis of representative individual K562 clones. (C) untransfected K562 cells. (A–D) Southern blots corresponding to the clones shown in the histogram panels above. All lanes are from the same gel (lines indicate where lanes were omitted) and were loaded with equivalent amounts of DNA. The unmarked lane between (C) and (D) is from another, irrelevant positive cell line.
- Figure S2. Quantitative chromatin immunoprecipitation (ChIP) analyses in nonerythroid cells (JPG, 15.5 KB)
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ChIP analyses of the 183bp region of the α-spectrin gene encoding exon 1′ and intron 1′ were performed with HeLa cell chromatin and antibodies against USF1 and USF2. A known USF binding site from a region of the DOLK gene was included as a positive control. Negative controls include GABRA6 and an unrelated region of the α-spectrin gene downstream of the 183bp region. The black bars represent USF1-precipitated chromatin, the gray bars represent USF2-precipitated chromatin, and the white bars represent IgG-precipitated chromatin.
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