title: GSM4672360: ChIP-seq H3K4me1 in human aortic valve interstitial cells; Homo sapiens; ChIP-Seq
library_construction_protocol: Human aortic valve interstitial cells were isolated from control non-mineralized aortic valves obtained from patients undergoing heart transplantation. The protocol was approved by the local ethical committee and informed consent was obtained from the subjects. Aortic leaflets were cut into pieces and incubated in 0.3% type I collagenase (Invitrogen, Thermo Fisher Scientific, ON, Canada) at 37°C for 45 minutes, then filtered through a 70 µm mesh, centrifuged for 5 minutes at 1,500 rpm and resuspended in complete media (DMEM, 10% FBS with L-glutamine and sodium pyruvate). Cells were used between passages 3 to 7. Nuclear extracts from VICs were immunoprecipitated with antibodies against H3K4me3, H3K4me1 (Cell Signaling Technology, New England Biolabs, ON, Canada) or H3K27Ac (Abcam, ON, Canada) pre-bound to protein G Dynabeads (Life Technologies, Thermo Fisher Scientific, ON, Canada). DNA was purified using DNA clean up & concentrator kit (Zymo Research, Cedarlane, Canada). Libraries were constructed using the QIAseq Ultralow Input Library Kit (Qiagen, ON, Canada) according to manufacturer's instructions. Sequencing was performed on an Illumina HiSeq4000 (UCSD IGM Genomics Facility, CA, USA).
identifiers.namespace: GEO
identifiers.id: GSM4672360