primary antibodies against mef2c  (ImmunoWay Biotechnology Company)

Journal: PLoS ONE

Article Title: Mef2c Regulates Transcription of the Extracellular Matrix Protein Cartilage Link Protein 1 in the Developing Murine Heart

doi: 10.1371/journal.pone.0057073

Figure Lengend Snippet: Crtl1 mRNA, Crtl1 protein, and Mef2c protein expression in the ventricular endocardium at ED10.5. (A) Crtl1 mRNA (blue staining, white arrow) is synthesized by ventricular endocardial cells at ED10.5. (B) Crtl1 protein expression (green) is observed in the ventricular cardiac jelly between the endocardial and myocardial cell layers. Mef2c protein expression (red) is observed in the nuclei of both the ventricular myocardium and ventricular endocardium (white arrows indicate endocardial Mef2c expression). endo = endocardium, myo = myocardium, LV = left ventricle.

Article Snippet: Eluted protein was run on a 4–20% Tris-glycine gel (Invitrogen, #EC6025), then subsequently transferred to a nitrocellulose membrane (Invitrogen, #LC2001), blocked in 5% dry milk/1%TBST, and probed with primary antibody against Mef2c (Santa Cruz, sc-13266).

Techniques: Expressing, Staining, Synthesized

Journal: PLoS ONE

Article Title: Mef2c Regulates Transcription of the Extracellular Matrix Protein Cartilage Link Protein 1 in the Developing Murine Heart

doi: 10.1371/journal.pone.0057073

Figure Lengend Snippet: (A) Atrioventricular junction in an H&E stained ED14.5 specimen. (B) Immunofluorescent staining of Crtl1 (green) shows Crtl1 is expressed in the ventricular aspect of the leaflets of the mitral valve at ED14.5, Mef2c (red, panel B) is also expressed throughout the leaflets of the mitral valve. (C) Atrioventricular junction in an H&E stained ED17.5 specimen. (D) Crtl1 is expressed sub-endocardially on the atrial aspects of the mitral valve leaflets at ED17.5 and Mef2c (red, panel F) is expressed in both the mesenchyme and endocardial lining of the leaflets, colocalizing with Crtl1 protein expression (green, panels D).

Article Snippet: Eluted protein was run on a 4–20% Tris-glycine gel (Invitrogen, #EC6025), then subsequently transferred to a nitrocellulose membrane (Invitrogen, #LC2001), blocked in 5% dry milk/1%TBST, and probed with primary antibody against Mef2c (Santa Cruz, sc-13266).

Techniques: Staining, Expressing

Journal: PLoS ONE

Article Title: Mef2c Regulates Transcription of the Extracellular Matrix Protein Cartilage Link Protein 1 in the Developing Murine Heart

doi: 10.1371/journal.pone.0057073

Figure Lengend Snippet: (A) Immunoblot for Mef2c protein. Lane 1, Mef2c binding to control Mef2 consensus sites from the muscle creatine kinase gene. Lane 2, negative control oligo with mutated Mef2 sites fails to bind to Mef2c. Lanes 3 and 5, Mef2c interacts with the Mef2 consensus sites in the Crtl1 promoter region at −913 to −923 (Site 1, Lane 3) and at −698 to −707 (Site 2, Lane 5). Lanes 4 and 6, mutation of the Mef2 consensus sites in the Crtl1 promoter region blocks Mef2 binding at both consensus sites. (B) Mef2c and Sox9 bind the Crtl1 Promoter in vivo . Chromatin Immunoprecipitation (ChIP) was performed using embryonic hearts stage ED10.5–11.0. PCR of the DNA input, Mef2c immunoprecipitate, and IgG control precipitate was performed using primers flanking both Mef2 consensus sites of the Crtl1 promoter. For the Sox9 ChIP, the initial DNA input, Sox9 immunoprecipitate, and IgG control precipitate were PCR amplified using primers specific for the Sox9 consensus site on the Crtl1 promoter.

Article Snippet: Eluted protein was run on a 4–20% Tris-glycine gel (Invitrogen, #EC6025), then subsequently transferred to a nitrocellulose membrane (Invitrogen, #LC2001), blocked in 5% dry milk/1%TBST, and probed with primary antibody against Mef2c (Santa Cruz, sc-13266).

Techniques: Western Blot, Binding Assay, Control, Negative Control, Mutagenesis, In Vivo, Chromatin Immunoprecipitation, Amplification

Journal: PLoS ONE

Article Title: Mef2c Regulates Transcription of the Extracellular Matrix Protein Cartilage Link Protein 1 in the Developing Murine Heart

doi: 10.1371/journal.pone.0057073

Figure Lengend Snippet: Fold change in luciferase activity driven by approximately 1 kb of the Crtl1 promoter in the pGL3 luciferase reporter vector was assayed in fetal chicken VICs (A and B) and in NIH3T3 cells (C and D). In fetal chicken VICs (A) and NIH3T3 cells (C), Crtl1 promoter activity was significantly increased with increasing concentrations of Mef2c. (B) Crtl1 promoter activity in the presence of 100 ng Mef2c with the addition of 200 ng of the Mef2-Engrailed dominant negative expression construct resulted in an approximately 30% reduction in Crtl1 reporter activity. (D) Mutations were introduced into the Crtl1 promoter construct at Mef2 Site 1 and Mef2 Site 2 (Crtl1-Mutant 1 and Crtl1-Mutant 2 respectively). Crtl1-Mutant 1 results in an approximately 30% reduction in Crtl1 promoter activation in the presence of 100 ng of Mef2c and Crtl1-Mutant 2 results in an approximately 50% reduction of Crtl1 activity.(*p<0.05, #p<0.1).

Article Snippet: Eluted protein was run on a 4–20% Tris-glycine gel (Invitrogen, #EC6025), then subsequently transferred to a nitrocellulose membrane (Invitrogen, #LC2001), blocked in 5% dry milk/1%TBST, and probed with primary antibody against Mef2c (Santa Cruz, sc-13266).

Techniques: Luciferase, Activity Assay, Plasmid Preparation, Dominant Negative Mutation, Expressing, Construct, Mutagenesis, Activation Assay