2025-06-01
ENA first public
ERC000011
ENA-CHECKLIST
Gehring Lab Institute for Genetics  University of Cologne
INSDC center name
public
INSDC status
ERS22945692
SRA accession
E-MTAB-14755:HFF_B_smgi
Submitter Id
ArrayExpress
broker name
TERT-HFF
cell_line
fibroblast
cell_type
not collected
collection date
human
common name
Protocols: Cells were harvested and lysed by adding 500 µl Invitrogen™ TRIzol™ Reagent (Invitrogen; Cat# 15596018) to each well. HFF (TERT-HFF; Ximbio Cat# 154459) and HUVEC (HUVEC/TERT 2; ATCC Cat# CRL-4053; RRID:CVCL_9Q53) cell lines were maintained at 37°C and 5% CO2 in a humidified incubator in (HFF:) DMEM + 10% FBS, + 1% MEM Sigma, Cat# M7145-100ml) or (HUVEC:) EGM Endothelial Cell Growth Medium BulletKit (Lonza, Cat# CC-3124) supplemented with 10% FBS and 20 µg/ml G418. 5x10^5 cells were seeded in 6-well plates one day before starting the SMG1i treatment. SMG1 inhibitor (SMG1i; DOI: 10.1016/j.bmcl.2012.08.107) dissolved in DMSO was added to a final concentration of 1 µM and cells were harvested 24 hours later. Total RNA was extracted using the Direct-zol RNA MiniPrep kit (Zymo Research; Cat# R2052) including the recommended DNase I treatment according to the manufacturer's instructions. poly(A) mRNA isolation and library preparation was carried out with 600 ng RNA as input using the NEBNext Poly (A) mRNA magnetic isolation module (NEB; Cat# E7490L) and NEBNext Ultra II Directional RNA library preparation kit (NEB; Cat# E7760L) according to the manufacturer's instructions.
description
neonate
developmental stage
normal
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
foreskin
organism part
Homo sapiens
scientific_name
male
sex
HFF_B_smgi
title