|
|
Blood, Vol. 108, Issue 12, 3736-3738, December 1, 2006
Acetylation of GATA-1 is required for chromatin occupancy
Blood Lamonica et al.
108: 3736
Supplemental materials for: Lamonica et al, Vol 108, Issue 12, 3736-3738
Files in this Data Supplement:
- Document 1. Supplemental methods (PDF, 72.9 KB)
- Figure S1. Acetylation of GATA-1 does not affect protein stability or nuclear localization (PDF, 547 KB) -
(A) Pools of cells infected with GATA-1–ER (G1) and GATA-1(NC)–ER (NC) express equal amounts of protein. Nuclear extracts (30 µg) from cells treated with tamoxifen were analyzed by Western blot with anti–GATA-1 antibodies. (B) Confocal immunofluorescence microscopy with antibodies against GATA-1 revealed comparable staining intensities and nuclear localization of GATA-1–ER and GATA-1(NC)–ER. As control, cells were stained with DAPI.
- Figure S2. Dramatic loss of gene activation and repression by GATA-1(NC)–ER (PDF, 20.7 KB) -
Real-time RT-PCR analysis of indicated genes. Signals were normalized to GAPDH and plotted as fold-activation (A) and repression (B), respectively.
- Figure S3. EMSA as described in the legend to Figure 1 (PDF, 1.31 MB) -
Probes contained TGATAG from HS3 (left), AGATAA from the Gata2 gene (middle), and AGATAG from the Gata2 gene (right). Note that, although absolute binding intensities varied among probes, GATA-1–ER and GATA-1(NC)–ER bound with similar avidity. Anti–GATA-1 and anti-ER antibodies were added where indicated.
- Figure S4. ChIP assays as described in the legend to Figure 2 using anti–GATA-1 (αG-1) and anti-ER (αER) antibodies, and control rat and mouse IgG (rIG and mIG) (PDF, 105 KB) -
Real-time PCR was used to analyze occupancy at natural Gata1 target sites, including the promoters of the  -major (A), Fog1 (C), Band3 (D), Ahsp (E), and Eklf (F) genes; HS3 of the -globin LCR (B); and the Gata2 (G) and c-Kit (H) genes. As negative control, a region upstream of the Fog1 locus was analyzed in parallel (C, left). Error bars indicate SD.
|
|
