Journal:

Article Title: A mutation in the translation initiation codon of Gata-1 disrupts megakaryocyte maturation and causes thrombocytopenia

doi: 10.1073/pnas.0606439103

Figure Lengend Snippet: The Plt13 mutation affects the translation initiation codon of Gata-1. (A) The PLT13 founder (G1, arrow) was identified because of a low platelet count (495 × 106 per milliliter) and bred to test the heritability of the phenotype. G2 mice that displayed a reduced platelet count (filled circles) were selected for breeding, and data from their offspring (G3) were separated by sex to demonstrate that the phenotype was X-linked. Platelet counts from G3 mice were tabulated (mean ± SD), and mice that had a platelet count <800 × 106 per milliliter were considered affected. (B) DNA sequence from exon 2 of Gata-1 is shown for wild-type (Gata-1+/+) and affected (Gata-1Plt13/+ and Gata-1Plt13/Y) animals. The translation initiation codon of Gata-1 is underlined. (C) 293T cells were transfected with expression constructs for wild-type Gata-1 (WT), Gata-1Plt13 (PLT13), or vector alone (V). Lysates were separated by SDS/PAGE, and a Western blot was performed with an antibody that recognizes the C terminus of Gata-1. Filled circles mark full-length Gata-1, and an open circle marks truncated Gata-1.

Article Snippet: Lysates prepared in KALB buffer were immunoprecipitated overnight with biotinylated goat anti-human GATA-1 (R & D Systems, Minneapolis, MN).

Techniques: Mutagenesis, Sequencing, Transfection, Expressing, Construct, Plasmid Preparation, SDS Page, Western Blot

Journal:

Article Title: A mutation in the translation initiation codon of Gata-1 disrupts megakaryocyte maturation and causes thrombocytopenia

doi: 10.1073/pnas.0606439103

Figure Lengend Snippet: Analysis of Gata-1 protein expression in megakaryocytes. Spleen sections were stained with antibodies that recognize the N terminus (in cyan) and the C terminus (in red) of Gata-1. CD41 staining was used to identify splenic megakaryocytes (in green). (A) In wild-type mice megakaryocytic nuclei stain with both N- and C-terminal antibodies. (B) There is a significant accumulation of megakaryocytes in Gata-1Plt13/+ mice, and many of these appear negative for Gata-1 staining (arrows).

Article Snippet: Lysates prepared in KALB buffer were immunoprecipitated overnight with biotinylated goat anti-human GATA-1 (R & D Systems, Minneapolis, MN).

Techniques: Expressing, Staining

Journal:

Article Title: A mutation in the translation initiation codon of Gata-1 disrupts megakaryocyte maturation and causes thrombocytopenia

doi: 10.1073/pnas.0606439103

Figure Lengend Snippet: Cell lines derived from Gata-1Plt13/+ mice have a mast cell phenotype. (A) Cells from a continuous mast cell line derived from a blast-like colony from Gata-1Plt13/+ bone marrow were cytocentrifuged and stained with May–Grunwald–Giemsa. (B) RT-PCR was performed on RNA extracted from continuous cell lines and from bone marrow (BM) with primers that flank the site of the Plt13 mutation (468-bp product). Digestion with NcoI cleaves the wild-type PCR product, resulting in two fragments (381 bp and 87 bp), but not the product from the Gata-1Plt13 allele. (C) Lysates from cell lines were immunoprecipitated with an antibody that binds to the C terminus of Gata-1 (+) or a control IgG from goat (−). Western blotting with the C-terminal M20 antibody demonstrated that Gata-1s protein (open circle) was absent in mast cell lines from Gata-1Plt13/+ mice, whereas the protein was detectable in lysates from transfected 293T cells. IgG heavy chain is marked by an arrow.

Article Snippet: Lysates prepared in KALB buffer were immunoprecipitated overnight with biotinylated goat anti-human GATA-1 (R & D Systems, Minneapolis, MN).

Techniques: Derivative Assay, Staining, Reverse Transcription Polymerase Chain Reaction, Mutagenesis, Immunoprecipitation, Western Blot, Transfection

Journal: Stem cells (Dayton, Ohio)

Article Title: β-globin-expressing definitive erythroid progenitor cells generated from embryonic and induced pluripotent stem cell-derived sacs

doi: 10.1002/stem.2335

Figure Lengend Snippet: (A–E). We analyzed RNA expression of several erythroid-related genes (BCL11a (A), cMYB (B), GATA1 (C), GATA2 (D), and KLF1 (E)) after 13-day erythroid differentiation (day 30) following CD34 or GPA selection from ES sac at day 15. We observed higher BCL11a expression, higher cMYB expression, lower GATA1 expression, lower GATA2 expression, and lower KLF1 expression in the CD34+ or GPA− cell fraction, compared to the CD34− or GPA+ cell fraction.

Article Snippet: We used TaqMan® Gene Expression Assays (Life Technologies) to evaluate RNA expression levels for cMYB (Hs00920556_m1), GATA1 (Hs01085823_m1), GATA2 (Hs00231119_m1), and KLF1 (Hs00610592_m1).

Techniques: RNA Expression, Selection, Expressing

Journal: International journal of oncology

Article Title: GATA-binding protein 1 is a novel transcription regulator of peroxiredoxin 5 in human breast cancer cells.

doi: 10.3892/ijo.2011.1236

Figure Lengend Snippet: Figure 2. GATA1-mediated modulation of both gene expression and promoter activity of Prx5. (A) GATA1 siRNA (13 nM) was transfected into MDA-MB- 231 cells, which have a high level of endogenous GATA1 expression, and Prx5 expression in these cells was confirmed using real-time quantitative RT-PCR and compared to those treated with vehicle control (‘MDA-MB-231 con’ in the figure). GAPDH was used as a control for normalization. (B) MDA-MB-231 cells were seeded in a 6-well plate. Twenty-four hours after transfection with either pGL3-F500 alone, a combination of pGL3-F500 and GATA1 siRNA (30 nM), or pGL3-F500 containing a mutation in the GATA1 binding site (pGL3-F500-GATA1 mt), the cells were harvested and their luciferase activity was determined. (C) pGL3 vectors containing various portions of the Prx5 promoter region were transfected into MCF7s cells and luciferase activities were measured as described in (B). Figure 3. Direct binding of GATA1 to the Prx5 promoter in cell lysates and cultured breast cancer cells. (A) A ChIP assay was performed in MDA-MB- 231 cells, which have high endogenous GATA1 expression levels, and in MCF7s cells using the antibodies and primer pairs described in ‘Materials and methods’. Binding affinity was measured by PCR. Binding affinity of mouse normal IgG was used as a negative control. An arrow indicates the PCR product corresponding to GATA1 binding site in the Prx5 promoter region. (B) EMSA was carried out in MDA-MB-231 cells using radiolabeled oligo nucleotides containing the wild-type or mutated sequences corresponding to the GATA1 binding site of the Prx5 promoter. Unlabeled oligonucleotides containing either wild-type or mutant GATA1 binding sites were used as a specific competitor. (C) EMSA was carried out in MCF7s cells using radio labeled oligonucleotides containing the GATA1 binding site of the Prx5 promoter.

Article Snippet: The membrane was probed with the primary antibody against each of the following protein; GATA1, Prx5, Bax, Bcl-2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and β-actin (sc-15, Sigma-Aldrich).

Techniques: Gene Expression, Activity Assay, Transfection, Expressing, Quantitative RT-PCR, Control, Mutagenesis, Binding Assay, Luciferase, Cell Culture, Negative Control, Labeling

Journal: International journal of oncology

Article Title: GATA-binding protein 1 is a novel transcription regulator of peroxiredoxin 5 in human breast cancer cells.

doi: 10.3892/ijo.2011.1236

Figure Lengend Snippet: Figure 4. Protection of cells from hydrogen peroxide-induced apoptosis via Prx5 expression (A) Prx5 siRNA was transfected into MCF7-Adr cells, which have high levels of endogenous Prx5. An inset picture shows the relative expression of Prx5 in cells treated with siRNA or no siRNA (control). FACS analysis was performed to measure H2O2-induced apoptosis in MCF7-Adr cells treated with Prx5 siRNA. Cells not exposed to H2O2 were used as a control to confirm annexinV staining. (B) Mean fluorescence intensities in MCF7-Adr cells with high or low Prx5 expression produced similar FACS histogram patterns. (C) GATA1 siRNA was transfected to MDA-MB-231 cells and the expression of GATA1 was detected by RT-PCR compared to samples treated with no siRNA (control). FACS analysis was performed as described in (A). (D) Mean fluorescence intensity in MDA-MB-231 cells with high or low GATA1 expression produced similar FACS histogram patterns. All experiments were performed in duplicate.

Article Snippet: The membrane was probed with the primary antibody against each of the following protein; GATA1, Prx5, Bax, Bcl-2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and β-actin (sc-15, Sigma-Aldrich).

Techniques: Expressing, Transfection, Control, Staining, Fluorescence, Produced, Reverse Transcription Polymerase Chain Reaction

Journal: International journal of oncology

Article Title: GATA-binding protein 1 is a novel transcription regulator of peroxiredoxin 5 in human breast cancer cells.

doi: 10.3892/ijo.2011.1236

Figure Lengend Snippet: Figure 5. Modulation of apoptotic/anti-apoptotic proteins in response to knockdown of either Prx5 or GATA1 expression. Samples designated as ‘Non’ represent those treated with no siRNA. Control samples that were not chal lenged with H2O2 are expressed as ‘No treat’. (A) Prx5 siRNA was transfected into MCF7-Adr cells. The cells were treated with H2O2 to induce apoptosis 24 h after transfection. The expression of Bcl-2, an anti-apoptotic protein, and Bax, a pro-apoptotic protein, were detected by Western blotting. β-Actin was used as a loading control. (B) GATA1 siRNA was transfected into MDA-MB-231 cells. Protein levels were determined as described in (A).

Article Snippet: The membrane was probed with the primary antibody against each of the following protein; GATA1, Prx5, Bax, Bcl-2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and β-actin (sc-15, Sigma-Aldrich).

Techniques: Knockdown, Expressing, Control, Transfection, Western Blot

Journal: International journal of oncology

Article Title: GATA-binding protein 1 is a novel transcription regulator of peroxiredoxin 5 in human breast cancer cells.

doi: 10.3892/ijo.2011.1236

Figure Lengend Snippet: Figure 6. Regulation of Prx5 gene expression via GATA1 and its physiological relevance in breast cancer cells. GATA1 acts as a transcription factor by binding to its binding site in the Prx5 promoter region and regulating Prx5 expression in a negative manner. GATA1 binding to the promoter region leads to reduction of transcription of Prx5 gene and Prx5 protein expression. Cells under oxidative stress may undergo apoptosis due to loss of Prx5-mediated protection against apoptosis.

Article Snippet: The membrane was probed with the primary antibody against each of the following protein; GATA1, Prx5, Bax, Bcl-2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and β-actin (sc-15, Sigma-Aldrich).

Techniques: Gene Expression, Binding Assay, Expressing

Journal: Frontiers in Pharmacology

Article Title: Regulating mitochondrial oxidative phosphorylation and MAPK signaling: wedelolactone as a novel therapeutic for radiation-induced thrombocytopenia

doi: 10.3389/fphar.2025.1508215

Figure Lengend Snippet: MAPK signaling pathway is necessary for WED-induced megakaryocyte maturation. (A-F) Western blot analysis showing the expression levels of RAS, p-MEK, p-ERK, FOS, NF-E2, and GATA1 in cells exposed to WED (2.5, 5, and 10 μM). *p < 0.05, **p < 0.01, ***p < 0.001 vs. the control group. (G) Flow cytometry analysis, with histograms displaying the proportion of CD41 + CD61 + cells. Data are displayed as mean ± SD (n = 3). ***p < 0.001 vs. the WED group.

Article Snippet: Add primary antibody and incubate overnight, then wash three times with PBST (PBS with 2% Tween-20), the secondary antibody was added and incubated at 37°C for 1 h. Antibodies against RAS (Abmart, T56672S), p-MEK1/2 (CST, 9154S), MEK1/2 (CST, 8727 S), p-ERK1/2 (Abmart, TA1015S), ERK1/2 (Abmart, TA0155), p-AMPK (Abmart, T55608F), AMPK (Abmart, T55326F), NF-E2 (Proteintech, United States, 11089-1-AP), GATA1 (Proteintech, United States, 10917-2-AP), FOS (Proteintech, United States, 66590-1-Ig), GAPDH (Proteintech, United States, 10494-1-AP).

Techniques: Western Blot, Expressing, Control, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Regulating mitochondrial oxidative phosphorylation and MAPK signaling: wedelolactone as a novel therapeutic for radiation-induced thrombocytopenia

doi: 10.3389/fphar.2025.1508215

Figure Lengend Snippet: Schematic model showing the role of WED in regulating megakaryocyte differentiation and platelet generation. WED promotes megakaryocyte differentiation and platelet generation by activating the MAPK signaling pathway (RAS/MEK/ERK1/2) and modulating oxidative phosphorylation via the AMPK signaling pathway. Activation of MAPK signaling results in the upregulation of key hematopoietic transcription factors, including FOS, NF-E2, and GATA1, which are essential for megakaryocyte differentiation and maturation. This process ultimately leads to the restoration of platelet counts, as demonstrated in the RIT mice model. MEP: Megakaryocyte-erythroid progenitor.

Article Snippet: Add primary antibody and incubate overnight, then wash three times with PBST (PBS with 2% Tween-20), the secondary antibody was added and incubated at 37°C for 1 h. Antibodies against RAS (Abmart, T56672S), p-MEK1/2 (CST, 9154S), MEK1/2 (CST, 8727 S), p-ERK1/2 (Abmart, TA1015S), ERK1/2 (Abmart, TA0155), p-AMPK (Abmart, T55608F), AMPK (Abmart, T55326F), NF-E2 (Proteintech, United States, 11089-1-AP), GATA1 (Proteintech, United States, 10917-2-AP), FOS (Proteintech, United States, 66590-1-Ig), GAPDH (Proteintech, United States, 10494-1-AP).

Techniques: Phospho-proteomics, Activation Assay

Journal: The Journal of Neuroscience

Article Title: Clonal Neural Stem Cells from Human Embryonic Stem Cell Colonies

doi: 10.1523/JNEUROSCI.3286-11.2012

Figure Lengend Snippet: TaqMan gene expression assays

Article Snippet: GATA1 , Hs00231112_m1.

Techniques: Gene Expression

Journal: The Journal of Neuroscience

Article Title: Clonal Neural Stem Cells from Human Embryonic Stem Cell Colonies

doi: 10.1523/JNEUROSCI.3286-11.2012

Figure Lengend Snippet: Clonal neurospheres grow in defined conditions over 4 weeks, express only neural markers, and could differentiate into neurons and glia. a, A floating clonal neurosphere at 28 d, grown from a single cell from an hESC colony. b, A comparison of mRNA transcript levels in hESCs versus primary neurospheres by Q-PCR revealed that clonal NSC colonies have an entirely neural phenotype: they express neural markers (NES, SOX1, PAX6, TUBB3) but not pluripotency genes (OCT4, LIN28), endodermal genes (FOXA2, HNF4A, GATA4), or mesodermal genes (GATA1, T), n = 4. c–g, Differentiation potential was assessed by plating single neurospheres on matrigel-coated plates in SFM with 1% FCS for 14 d. Differentiated neurospheres produced TUBB3+ (c) and MAP2+ (g) neurons (shown here growing out of a whole sphere), GFAP+ (e) astrocytes, and O4+ (f) oligodendrocytes, while many cells still expressed the neural precursor marker NES (d). All nuclei were counterstained with Hoechst stain (blue). h, Quantification of marker proteins by ICC show that differentiated cell types consisted of neural precursors (45.9% ± 6.9% NES+), neurons (37.9% ± 8.3% TUBB3+, and 15.2% ± 4.2% MAP2 +), astrocytes (34.7% ± 4.2% GFAP+), and oligodendrocytes (5.8% ± 1.9% O4+). i, A population of differentiating cells colocalize neural precursor marker NES, and astrocyte marker GFAP. Cells were double labeled with GFAP (red) and NES (green) to reveal a population of differentiating cells positive for both proteins (yellow). This overlapping expression accounts for combined marker expression in h being >100%. j, k, Differentiation to 5 weeks produced GABAergic (j) and dopaminergic (k) neurons. Data are represented as mean ± SEM; for Q-PCR, the highest expressing sample (value of “1”) was used as the calibrator.

Article Snippet: GATA1 , Hs00231112_m1.

Techniques: Comparison, Produced, Marker, Staining, Labeling, Expressing

Journal: bioRxiv

Article Title: Emergence of the erythroid lineage from multipotent hematopoiesis

doi: 10.1101/261941

Figure Lengend Snippet: a qRT-PCR for expression of established erythroid regulators in sorted P1 to P5 subsets. Expression of each gene is normalized to p-actin and is the mean of at least two experiments. b Western blot analysis for GATA1 expression in sorted P1 (subdivided into CD71 med and CD71 hi subsets), P2, and EryA (CD71+Ter119+FSC hi cells, representative of ETD). c Density of FACS subsets P1 to P5 along the erythroid trajectory. Single-cell transcriptomes from each subset were mapped to their most similar counterparts in the Kit+ data . Shown here is the fraction of mapped cells, following smoothing with a Gaussian kernel. d Distribution of CD71 expression in P1 and P2 cells immediately following sorting (gray) and after 24 hours of in vitro differentiation (lavender). e Dynamically varying genes along the MPP to erythroid axis were clustered based on their behavior across three transition points. At each transition, gene expression is either increased, decreased, or unchanged, giving a total of 27 potential dynamic patterns across all 3 transitions, shown in red. The number of genes corresponding to each dynamic pattern is noted, and individual gene Z-score normalized expression traces are shown in black. Selected clusters shown in are marked with an asterisk and a representative gene.

Article Snippet: The following TagMan MGB probes were used: Mst1r (Mm00436382_m1), Ryk (Mm01238551_m1), IL17ra (Mm00434214_m1), Mt2 (Mm00809556_s1), Slc26a1 (Mm01198850_m1), Slc4a1 (Mm00441492_m1), Trib2 (Mm00454876_m1), Cd34 (Mm00519283_m1), Meis1 (Mm00487664_m1), Hpn (Mm01152654_m1), Pf4 (Mm00451315_g1), Dntt, (Mm00493500_m1), Ms4a2 (Mm00442778_m1), Elane (Mm00469310_m1), S100a9 (Mm00656925_m1), F13a1 (Mm00472334_m1), Egr1(Mm00656724_m1), Apoe (Mm01307193_g1), Ldb1 (Mm00440156_m1), Zfpm1 (Mm00494336_m1), Tfrc (Mm00441941_m1), Hbb-b1 (Mm01611268_g1), Alas2 (Mm01260713_m1), Band3 (Mm01245920_g1), Nfe2 (Mm00801891_m1), Gata1 (Mm01352636_m1), Gata2 (Mm00492300_m1), Klf1 (Mm00516096_m1) and PU1/Sfpi1 (Mm00488393_m1).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, In Vitro

Journal: Nature communications

Article Title: RASSF1A uncouples Wnt from Hippo signalling and promotes YAP mediated differentiation via p73.

doi: 10.1038/s41467-017-02786-5

Figure Lengend Snippet: Fig. 1 Epigenetic activation of Rassf1A drives ESC differentiation via GATA1. a RASSF1A and Pou5f1/Oct4 mRNA levels in undifferentiated and differentiating human ESC lines H1 and H9. Data obtained from published GSE54186 data set. b Rassf1A mRNA levels increase upon LIF withdrawal-mediated differentiation of mouse ESC (mESC) lines V6.5 and J1. Data obtained from published GSE3749 and GDS2672 data sets. c Rassf1A mRNA (bars) and protein (blots) levels from undifferentiated (+LIF) versus differentiated (-LIF) E14Tg2a mESC. See also Supplementary Fig. 1b and Supplementary Movie 1. d Rassf1A CpG island promoter methylation (assessed by qPCR for methylated/unmethylated DNA ratio) and e H3K4me3 levels in pluripotent (+LIF) versus differentiated (-LIF) mESC. See also Supplementary Fig. 1a. f ENOCDE transcription factor ChIP data showing specific binding of GATA1 on the Rassf1A promoter of differentiated hESC and GATA1 ChIP on the Rassf1A promoter of pluripotent (+LIF) versus differentiated (-LIF) ESC. g qPCR for Rassf1A and Oct4 mRNA levels in response to siRNA-mediated Gata1 silencing in the presence or absence of LIF in ESC. Western blotting and qPCR indicate the level of Gata1 KD. See also Supplementary Figs 1d–e, 2c and 5d. h Representative ESC colonies from indicated conditions subject to LIF withdrawal assay. Scale bars: 25–50 μm. **P < 0.01 of Student’s t-test. Error bars indicate s.e.m. Data shown are representative of at least three independent experiments

Article Snippet: ESCs transfected with pLKO.1 puro were cultured in the presence of 1.75 μg/mL of puromycin for 2 days. pcDNA3 GATA1 was a gift from Licio Collavin & Giannino Del Sal (Addgene plasmid # 85693).

Techniques: Activation Assay, Methylation, Binding Assay, Western Blot

Journal: Journal of Clinical Investigation

Article Title: Ribonuclease inhibitor 1 regulates erythropoiesis by controlling GATA1 translation

doi: 10.1172/jci94956

Figure Lengend Snippet: Figure 6. GATA1 overexpression rescues erythroid phenotype in Rnh1- deficient yolk sac cells. (A) Total number of colonies observed in control and GATA1- infected Rnh1-deficient yolk sac cells that were cultured for 7 days in methyl cellulose medium (n = 3). Data are shown as mean ± SD. P values were determined by 2-tailed t test. (B) Cytospin images of erythroid cells derived from methyl cellulose colonies of control and GATA1-infected cells. More mature erythroid cells were observed in GATA1-infected cells. Original magnification ×400.

Article Snippet: Using these whole cell lysates, immunoprecipitation was performed with GATA1 antibody (catalog 4591, Cell Signaling Technology) bound to Dynabeads Protein G (10003D, Life Technologies) for 3 hours with rotation at 4°C.

Techniques: Over Expression, Control, Infection, Cell Culture, Derivative Assay

Journal: Journal of Clinical Investigation

Article Title: Ribonuclease inhibitor 1 regulates erythropoiesis by controlling GATA1 translation

doi: 10.1172/jci94956

Figure Lengend Snippet: Figure 7. RNH1 deficiency decreases erythroid differ- entiation and GATA1 protein levels in K562 cell. (A) Total protein lysates of WT and RNH1-KO K562 cells were analyzed by Western blot with the indicated antibodies. K562 cells were positive for BCR-ABL oncogene. Blots are representative of 3 indepen- dent experiments. (B) qRT- PCR analysis for GATA1 mRNA levels in K562 cells, normal- ized to 18S rRNA expression. Data are shown as mean ± SD and are representative of 3 independent experiments. (C) Number of benzidine-positive K562 cells. Data are shown as mean ± SD and are represen- tative of 3 independent exper- iments. (D) K562 cells were treated with hemin (25 μM) for 3 days and analyzed for the presence of benzidine-positive cells. Data are shown as mean ± SD and are representative of 3 independent experiments. Original magnification ×200. (E) Polysome profiles for WT and RNH1-KO K562 cells. Data are representative of 3 independent experiments. (F) qRT-PCR analysis for indicated mRNAs in monosome and polysome fractions derived from WT and RNH1-KO K562 cells, normalized to 18S rRNA expression. Data are shown as mean ± SD and are repre- sentative of 3 independent experiments. P values were determined by 2-tailed t test.

Article Snippet: Using these whole cell lysates, immunoprecipitation was performed with GATA1 antibody (catalog 4591, Cell Signaling Technology) bound to Dynabeads Protein G (10003D, Life Technologies) for 3 hours with rotation at 4°C.

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Derivative Assay

Journal: Journal of Clinical Investigation

Article Title: Ribonuclease inhibitor 1 regulates erythropoiesis by controlling GATA1 translation

doi: 10.1172/jci94956

Figure Lengend Snippet: Figure 12. RNH1 is a ribosomal- associated protein that regulates erythropoiesis by controlling GATA1 translation. RNH1 is a ribosomal- associated protein involved in transla- tion of GATA1, which leads to efficient differentiation of progenitor cells to mature erythroid cells. Lack of RNH1 decreases GATA1 levels, which leads to an arrest in differentiation.

Article Snippet: Using these whole cell lysates, immunoprecipitation was performed with GATA1 antibody (catalog 4591, Cell Signaling Technology) bound to Dynabeads Protein G (10003D, Life Technologies) for 3 hours with rotation at 4°C.

Techniques: