Journal: Molecular and Cellular Biology
Article Title: Chromatin Architecture and Transcription Factor Binding Regulate Expression of Erythrocyte Membrane Protein Genes
doi: 10.1128/mcb.00777-09
Figure Lengend Snippet: FIG. 2. Representative integrated genome browser (IGB) views of RNAPII, H3Me3K4, GATA-1, and NF-E2 binding at the 5 end of the -spectrin gene (A to C) and the ERMAP gene locus (D to F). (A) IGB view of RNAPII binding at the 5 end of the -spectrin promoter in erythroid (K562) and nonerythroid (HeLa) cells. There is a peak of RNAPII binding at the TSS of the -spectrin gene in K562 cells but not in HeLa cells, demonstrating cell type-specific binding of RNAPII. (B) IGB view of H3Me3K4 binding at the 5 end of the -spectrin gene in erythroid (K562) and nonerythroid (HeLa) cells. There is a peak of H3Me3K4 binding in K562 cells but not in HeLa cells, demonstrating cell type-specific enrichment for H3Me3K4. (C) IGB view of GATA-1 and NF-E2 binding at the 5 end of the -spectrin gene, demonstrating NF-E2 binding at the promoter and GATA-1 binding in intron 3. (D) IGB view of RNAPII binding at the ERMAP locus in erythroid (K562) and nonerythroid (HeLa) cells. Two ERMAP isoforms are shown. There is a peak of RNAPII at the TSS in both K562 and HeLa cells for one isoform and a peak of RNAPII in K562 cells only for the alternate isoform, demonstrating cell type-specific binding of RNAPII. (E) IGB view of H3Me3K4 binding at the ERMAP locus in erythroid (K562) and nonerythroid (HeLa) cells. The ERMAP isoform with the erythroid cell-specific RNAPII binding has greater enrichment for H3Me3K4 in K562 cells than in HeLa cells, demonstrating cell type-specific enrichment of H3Me3K4. (F) IGB view of GATA-1 and NF-E2 binding at the ERMAP locus, demonstrating NF-E2 binding at the promoter, in intron 1, and at the 3 adjacent region and GATA-1 binding in intron 1 and at the 3 adjacent region of the ERMAP locus.
Article Snippet: Antibodies utilized for immunoprecipitation included H3Me3K4 (ab8580; Abcam), RNA polymerase II (RNAPII) (sc-899; Santa Cruz), GATA-1 (sc-265; Santa Cruz), and NF-E2 (sc-22827; Santa Cruz) antibodies, as well as nonspecific rabbit immunoglobulin G (IgG) (sc-2091; Santa Cruz).
Techniques: Binding Assay