Project description:Low levels of Cdh2-BioID2 were expressed in mouse primary neonatal cardiomyocytes using an adenoviral expression system. Infected cells were treated with biotin for 24 hours before lysis. Biotinylated proteins were precipitated from lysates using streptavidin beads. Protein samples were in-gel trypsin digested and the resulting tryptic peptides were analyzed on an LTQ Orbitrap Velos.
Project description:The adenovirus carrying Mettl14 gene infects primary neonatal mouse cardiomyocytes, which is in contrast with the primary neonatal mouse cardiomyocytes infected with empty virus. The purpose is to explore the effect of Mettl14 on the global mRNA m6A modification level in primary neonatal rat cardiomyocytes.
Project description:To investigate the impact of mutant RIT1 expression in cardiac cells, we isolated neonatal cardiomyocytes from mice harbouring an engineered Rit1 locus with Cre recombinase-inducible expression of the pathogenic variant Rit1M90I. Upon isolation, cardiomyocytes were treated with adenoviruses encoding for Cre recombinase to induce expression of the Rit1M90I variant.
Project description:Low levels of Cdh2-BioID2 were expressed in mouse primary neonatal cardiomyocytes using an adenoviral expression system. Infected cells were treated with biotin for 24 hours before lysis. Biotinylated proteins were precipitated from lysates using streptavidin beads. Protein samples were in-gel trypsin digested and the resulting tryptic peptides were analyzed on an LTQ Orbitrap Velos.
Project description:To gain insight into the molecular regulation of human heart development, a detailed comparison of the mRNA and miRNA transcriptomes across differentiating human-induced pluripotent stem cell (hiPSC)–derived cardiomyocytes and biopsies from fetal, adult, and hypertensive human hearts was performed. Gene ontology analysis of the mRNA expression levels of the hiPSCs differentiating into cardiomyocytes revealed 3 distinct groups of genes: pluripotent specific, transitional cardiac specification, and mature cardiomyocyte specific. Hierarchical clustering of the mRNA data revealed that the transcriptome of hiPSC cardiomyocytes largely stabilizes 20 days after initiation of differentiation. Nevertheless, analysis of cells continuously cultured for 120 days indicated that the cardiomyocytes continued to mature toward a more adult-like gene expression pattern. Analysis of cardiomyocyte-specific miRNAs (miR-1, miR-133a/b, and miR-208a/b) revealed a miRNA pattern indicative of stem cell to cardiomyocyte specification. A biostatistitical approach integrated the miRNA and mRNA expression profiles revealing a cardiomyocyte differentiation miRNA network and identified putative mRNAs targeted by multiple miRNAs. Together, these data reveal the miRNA network in human heart development and support the notion that overlapping miRNA networks re-enforce transcriptional control during developmental specification. Comparison of mRNA expression profiling of differentiating human-induced pluripotent stem cell (hiPSC)–derived cardiomyocytes, biopsies from fetal, adult and hypertensive human hearts and primary cardiomyocytes
Project description:Dysregulation of miRs has been reported in a variety of cardiac diseases. In particular, it has reported that estrogen regulates a miRs network in female cardiac fibroblasts, thereby modulating a spectrum of genes involved in cardiac fibrosis and remodeling. However, the estrogen-responsive miRs in cardiomyocytes still remain to be elucidated. We used a microRNA microarray screening approach to address the miRs expression profiling in estrogen-treated cardiomyocytes.