WL3844
Protocols: Human TeloHAECs were grown in cell culture Cells were dounced in freshly prepared solution A ( 1% formaldehyde, 50mM Hepes-KOH, 100mM NaCl, 1mM EDTA, 0.5mM EGTA) and incubated for 20 mins at room temperature. The formaldehyde was then quenched with 1/20 volume of 2.5M glycine and incubated for 5 min. Tissue was then rinsed with ice-cold PBS twice and centrifuged at 4 C at 2500 x rcf for 5 min. Crosslinked tissue was then lysed using Chromatin Easy Shear Low SDS Kit (Diagenode #C01020013). Chromatin was sheared into 100–500 bp DNA fragments by sonication (Diagenode Bioruptor Pico) for 8 cycles of 30s ON and 30s OFF. Approximately 1.5% of chromatin was used for input DNA extraction. For each ChIP, chromatin lysates were combined with 10ug of anti-H3K27ac (Active Motif #39133), anti-H3K4me2 (Millipore # 07-030), H3K27me3 (Millipore #07-449), H3K36me3 (Abcam #ab9050) and H3K9me2 (Abcam #ab8898) antibodies and incubated overnight rotating at 4C.  For library preparation, all the ChIP DNA and input DNA were mixed with reagents from NEBNext Ultra II Library Prep Kit (NEBNext #E7645S) according to the manufacturer's protocol. This DNA was then amplified with barcoded primers for Illumina sequencing (NEBNext® Multiplex Oligos for Illumina® #E7335S). PCR amplifications were carried out as [98 °C 30 s, (98 °C 10 s, 65 °C 75s) ×9cycles, 65 °C 5 min, 4 °C hold]. The amplified and barcoded library was then purified and selected for 200–350-bp fragments using AMPure XP beads (BECKMAN COULTER #A63881)
ENA first public
2026-08-05
INSDC center name
The Hospital for Sick Children, Toronto Ontario, Canada The Ontario Institute for Cancer Research, Toronto, Ontario, Canada
INSDC status
public
Submitter Id
E-MTAB-17311:WL3844
broker name
ArrayExpress
cell type
aortic endothelial cell
collection date
not collected
common name
human
developmental stage
cell line
disease
normal
genotype
wild type genotype
geographic location (country and/or sea)
not collected
isolate
not applicable
obsolete_cell line
TeloHAEC
organism part
aorta endothelium
sample name
E-MTAB-17311:WL3844
scientific_name
Homo sapiens