title: 1248 + ET1 -1
organism: Mus musculus
description: 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.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-06-01
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-06-01
attributes.tag: External Id
attributes.value: SAMEA112752411
attributes.tag: INSDC center alias
attributes.value: UNIVERSITY OF EASTERN FINLAND
attributes.tag: INSDC center name
attributes.value: UNIVERSITY OF EASTERN FINLAND
attributes.tag: INSDC first public
attributes.value: 2023-06-01T00:15:37Z
attributes.tag: INSDC last update
attributes.value: 2023-06-01T00:15:37Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-12741:1248 + ET1 -1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: cardiac muscle cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: neonate
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: cardiac ventricle
attributes.tag: sample name
attributes.value: E-MTAB-12741:1248 + ET1 -1
attributes.tag: scientific_name
attributes.value: Mus musculus