Project description:We addressed the question of which protein kinases are expressed in cardiomyocytes. We assessed the changes during postnatal development, comparing profiles in rat neonatal ventricular cardiomyocytes (NVMs) with adult ventricular cardiomyocytes (AVMs). Neonatal and adult rat ventricular cardiomyocytes prepared according to established procedures (Marshall et al. PLoS ONE 2010 5(4):e10027; Fuller and Sugden, FEBs Lett. 1989 247:209-12; Rodrigues and Severson In Biochemical Techniques in the Heart (McNeill, J. H., Ed.) pp 101-115, CRC Press, New York.). mRNA expression profiles compared using Affymetrix rat genome 230 2.0 arrays.
Project description:miR-222 overexpression leads to promotion of proliferation and hypertrophy and inhibition of apoptosis in in primary neonatal rat ventricular cardiomyocytes (NRVMs). Isolated primary neonatal rat ventricular cardiomyocytes were plated in 6 cm BD Primaria tissue culture dishes. Transfection of microRNA precursors or scramble control (0.4 μM) was carried out using Lipofectamine RNAiMAX (Invitrogen) as recommended by the manufacturer. Forty-eight hours after transfection, RNAs from cultured cells and tissues were isolated with Tryzol (Invitrogen) following the manufactures’ manuals. Total RNA was harvested and submitted to the Dana-Farber Cancer Institute Molecular Diagnostics Laboratory for assay. These results revealed miR-222 regulated gene expression in primary neonatal rat ventricular cardiomyocytes.
Project description:miR-222 overexpression leads to promotion of proliferation and hypertrophy and inhibition of apoptosis in in primary neonatal rat ventricular cardiomyocytes (NRVMs). Isolated primary neonatal rat ventricular cardiomyocytes were plated in 6 cm BD Primaria tissue culture dishes. Transfection of microRNA precursors or scramble control (0.4 μM) was carried out using Lipofectamine RNAiMAX (Invitrogen) as recommended by the manufacturer. Forty-eight hours after transfection, RNAs from cultured cells and tissues were isolated with Tryzol (Invitrogen) following the manufacturesâ?? manuals. Total RNA was harvested and submitted to the Dana-Farber Cancer Institute Molecular Diagnostics Laboratory for assay. These results revealed miR-222 regulated gene expression in primary neonatal rat ventricular cardiomyocytes. Gene expression in NRVM cells tranfected with control scramble precursor (4 samples) or miR-222 precursor (4 smaples ) was evaluated using Affymetrix Rat Genome 230 v2.0.
Project description:Ten million neonatal rat ventricular cardiomyocytes were infected with adenovirus to overexpress NRF1 or LacZ (control). 48 hours after infection, cells were collected and lysed for TMT proteomic analysis.
Project description:We performed a set of sequencing experiments to identify the targets of Vascular endothelial zinc finger 1 (Vezf1) in neonatal rat cardiomyocytes. We compared a set of control samples to samples transfected with Vezf1 siRNA by high throughput sequencing. Vascular endothelial zinc finger 1 (Vezf1) siRNA was transfected into the neonatal rat ventricular cardiomyocytes for 24h and thereafter the cells were incubated in serum free medium for 48h.
Project description:Study on changes in gene expression in primary cultures of neonatal rat ventricular cardiomyocytes to electric stimulation. Through comparing non-stimulated, stimulated and blebbistatin supplemented and stimulated cultures we set out to identify the genes that are specifically activated by electric pulsing separate from cardiomyocyte contractions. After initial recovery phase, primary cultures of neonatal rat ventricular cardiomyocytes were cultured for 3 days without pulsing, with electric pulsing or with electric pulsing combined with blebbistatin. Per treatment: 3 arrays, with samples obtained from 3 separate series of cardiomyocyte isolation and culturing. Per array: sample prepared from pooled (1:1) RNA from duplicate experiments.
Project description:The goal of this experiment is to identify differentially expressed proteins in RBX2-deficient cardiomyocytes. Neonatal rat ventricular cardiomyocytes were transfected with indicated siRNAs and treated with or without CCCP for 6 hours. The collected cell lysates were processed for tandem mass tag labeling and quantitative proteomics analysis.
Project description:Using the highly sensitive RNA array, we assessed the effect of 11β-HSD1 and BVT.2733 on modulating RNAs expression in the neonatal rat ventricular cardiomyocytes.
