characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-06-01T00:15:37Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-06-01T00:15:37Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112752422
characteristics.characteristic: External Id
characteristics.text: UNIVERSITY OF EASTERN FINLAND
characteristics.characteristic: INSDC center name
characteristics.text: 2023-06-01T00:15:37Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-06-01T00:15:37Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14747112
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12741:DMSO - 3
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: cardiac muscle cell
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Samples of bulk RNA from cell cultures were collected from primary cell cultures of neonatal mouse ventricular cardiomyocytes by addition of the Macherey-Nagel Nucleospin lysis buffer prior to RNA isolation. Neonatal mouse hearts were removed from pups, and right and left neonatal mouse ventricles were separated from the atria using a scalpel and scissors in cold PBS. Following dissociation, isolated ventricular cardiomyocytes were cultured prior to experiments. Primary neonatal mouse ventricular cardiomyocytes (NMVCMs) were isolated using the neonatal heart dissociation kit (Miltentyi) and neonatal cardiomyocyte isolation kit (Milentyi) and plated to 12-well plates (200k cells/well). Cardiomyocytes were cultured for 24 hours in DMEM/F12 supplemented with 2.5 mg/mL bovine serum albumin, insulin-transferrin-selenium, sodium pyruvate, pen-strep, and T3 prior to ET-1 (Sigma) treatment and/or compound addition. Individual replicates represent different isolation batches, and cells were not cryopreserved at any stage prior to experiments. The test compound was diluted in DMSO, and 0.1% DMSO was used as control. DMSO or test compound 3i-1248 was added to cells at 1µM for 24 hours prior to sample collection. RNA isolation was performed using the Macherey-Nagel Nucleospin kit with on-column DNAse treatment. Library construction was performed based on multiplexing with Drop-Seq, using indexing oligonucleotides to multiplex samples, followed by cDNA synthesis and PCR amplification. PCR amplicons were tagged and libraries prepared using Illumina reagents.
characteristics.characteristic: description
characteristics.text: neonate
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: cardiac ventricle
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: DMSO - 3
characteristics.characteristic: title