Gata4

Manufactured by R&D Systems

GATA4 is a transcription factor that plays a crucial role in the development and regulation of the cardiovascular system. It is a member of the GATA family of zinc-finger transcription factors and is primarily expressed in cardiac and endodermal tissues.

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Lab products found in correlation

Facscalibur, by BD (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Axiovert 200, by Zeiss (1 mentions) Cd105, by Abcam (1 mentions) Mouse igg1 apc, by Miltenyi Biotec (1 mentions) Cardiac troponin 1, by Abcam (1 mentions) Mouse igg1 fitc, by R&D Systems (1 mentions) Sarcomeric α actinin, by Merck Group (1 mentions) Sm22 alpha, by Abcam (1 mentions) Calponin, by Abcam (1 mentions) Texas red affinipure donkey anti mouse igg, by Jackson ImmunoResearch (1 mentions) Laminin, by Abcam (1 mentions) Fitc affinipure donkey anti rabbit igg h l, by Jackson ImmunoResearch (1 mentions) Alexa fluor 594 donkey anti goat, by Thermo Fisher Scientific (1 mentions)

3 protocols using gata4

Characterization of Cardiac Progenitor Cells

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CDCs were dissociated to single cell suspension by incubation with 0.05% Trypsin-EDTA and resuspended in DMEM/F12 with HEPES (Gibco) supplemented with 10% FBS (FACS Staining Medium). They were incubated with conjugated antibody at 1:10 dilution for 1 h at 4°C prior flow cytometry analysis (BD FACSCalibur). Antibodies used include CD105 (mouse monoclonal, Abcam), CD90 (mouse monocolonal, BioLegend), CD45 (mouse monocolonal, AbD Serotec), mouse IgG1-FITC (R&D Systems), and mouse IgG1-APC (Miltenyi Biotec). For immunocytochemistry, cells were fixed with 4% paraformaldehyde and immunostained with primary antibodies GATA4 (goat polyclonal, R&D Systems), Nkx2.5 (goat polyclonal, R&D Systems), and cardiac troponin I (mouse monoclonal, Abcam). Primary antibodies were used at a 1:100 dilution. Alexa Fluor conjugated 488 or 555 donkey anti-mouse, goat or rabbit secondary antibodies (Molecular Probes) were added at a dilution of 1:500 for 1 hour at room temperature. Fixed cultures were counterstained with DAPI (Sigma). Images were obtained using a Zeiss Axiovert 200 inverted fluorescence microscope.

Lang J.K., Young R.F., Ashraf H, & Canty JM J.r. (2016). Inhibiting Extracellular Vesicle Release from Human Cardiosphere Derived Cells with Lentiviral Knockdown of nSMase2 Differentially Effects Proliferation and Apoptosis in Cardiomyocytes, Fibroblasts and Endothelial Cells In Vitro. PLoS ONE, 11(11), e0165926.

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Immunostaining of Pluripotent and Cardiac Cells

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The iPS cells and the cells at day 22–28 of iPS EB differentiation were fixed with 4% paraformaldehyde, and stained following a standard immunostaining procedure. Briefly, cells were fixed in 4% formaldehyde solution in phosphate-buffered saline (PBS), permeated with 5% bovine serum albumin (BSA) plus 0.1 Triton X100, and blocked with 5% BSA plus 0.01% tween 20. The primary and secondary antibodies were diluted in blocking solution. The primary antibodies against mouse Nanog (cat# ab80892, Abcam), Sox2 (cat# ab5603, Millipore), Oct4 (cat# sc-5279, Santa Cruz), Connex43 (cat# C6219, Sigma);α-sarcomeric actinin (cat# A7732, Sigma); MHC (ab15, Abcam); cTnT (ab8295, Abcam); Laminin (cat# ab11575, Abcam); SM22 alpha (cat# ab14106, Abcam); alpha smooth muscle Actin (cat# ab5694,Abcam); Calponin (cat# ab46794, Abcam); GATA4 (cat# AF2606, R&D system); and NKx2.5 (cat# AF2444, R&D system) were used at a 1:200 dilution. Secondary antibodies, Texas Red AffiniPure Donkey Anti-Mouse IgG (cat#715-075-150, Jackson ImmunoResearch), Texas Red AffiniPure Donkey Anti-Goat IgG (H+L) (cat# 705-075-003, Jackson ImmunoResearch), FITC AffiniPure Donkey Anti Rabbit IgG (H+L) (cat# 711-095-152, Jackson ImmunoResearch), and Texas Red AffiniPure Donkey Anti- Rabbit IgG (H+L) (Cat#711-075-152, Jackson ImmunoResearch) were used at a 1:200 dilution.

Wang H., Xi Y., Zheng Y., Wang X, & Cooney A.J. (2016). Generation of Electrophysiologically Functional Cardiomyocytes from Mouse Induced Pluripotent Stem Cells. Stem cell research, 16(2), 522-530.

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Immunocytochemistry Visualization of GATA4

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Standard immunocytochemistry methods were used^{49 (link)}. Briefly, cells were fixed with 4% PFA/1X PBS, permeabilized with 0.5% Triton X-100/1X PBS, blocked with 3% BSA/1X PBS, and incubated with antibodies (GATA4, R&D Systems, AF2606; Alexa-Fluor 594 Donkey anti-goat, Thermo Fisher) in 1% BSA/1X PBS. DAPI was used to visualize nuclei. Images were taken with Nikon Eclipse TE300 fluorescent microscope equipped with SPOT RT3 camera with 20X objective. Images were assembled into figures using Adobe Photoshop and Illustrator. Images from control and experimental samples were processed identically.

Stavniichuk R., DeLaForest A., Thompson C.A., Miller J., Souza R.F, & Battle M.A. (2021). GATA4 blocks squamous epithelial cell gene expression in human esophageal squamous cells. Scientific Reports, 11, 3206.

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